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WO1999013943A1 - Ensemble a ultrasons destine a etre utilise avec des medicaments actives par la lumiere - Google Patents

Ensemble a ultrasons destine a etre utilise avec des medicaments actives par la lumiere Download PDF

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Publication number
WO1999013943A1
WO1999013943A1 PCT/US1998/019797 US9819797W WO9913943A1 WO 1999013943 A1 WO1999013943 A1 WO 1999013943A1 US 9819797 W US9819797 W US 9819797W WO 9913943 A1 WO9913943 A1 WO 9913943A1
Authority
WO
WIPO (PCT)
Prior art keywords
light activated
ultrasound
tissue site
catheter
drag
Prior art date
Application number
PCT/US1998/019797
Other languages
English (en)
Inventor
Katsuro Tachibana
Shunro Tachibana
James R. Anderson
Gary Lichttenegger
Original Assignee
Ekos Corporation
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from JP25581497A external-priority patent/JP4791616B2/ja
Priority claimed from US09/129,980 external-priority patent/US6210356B1/en
Application filed by Ekos Corporation filed Critical Ekos Corporation
Priority to AU95005/98A priority Critical patent/AU9500598A/en
Publication of WO1999013943A1 publication Critical patent/WO1999013943A1/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M37/00—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin
    • A61M37/0092—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin using ultrasonic, sonic or infrasonic vibrations, e.g. phonophoresis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0047—Sonopheresis, i.e. ultrasonically-enhanced transdermal delivery, electroporation of a pharmacologically active agent
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B17/00—Surgical instruments, devices or methods
    • A61B17/22—Implements for squeezing-off ulcers or the like on inner organs of the body; Implements for scraping-out cavities of body organs, e.g. bones; for invasive removal or destruction of calculus using mechanical vibrations; for removing obstructions in blood vessels, not otherwise provided for
    • A61B17/22004—Implements for squeezing-off ulcers or the like on inner organs of the body; Implements for scraping-out cavities of body organs, e.g. bones; for invasive removal or destruction of calculus using mechanical vibrations; for removing obstructions in blood vessels, not otherwise provided for using mechanical vibrations, e.g. ultrasonic shock waves
    • A61B17/22012—Implements for squeezing-off ulcers or the like on inner organs of the body; Implements for scraping-out cavities of body organs, e.g. bones; for invasive removal or destruction of calculus using mechanical vibrations; for removing obstructions in blood vessels, not otherwise provided for using mechanical vibrations, e.g. ultrasonic shock waves in direct contact with, or very close to, the obstruction or concrement
    • A61B17/2202—Implements for squeezing-off ulcers or the like on inner organs of the body; Implements for scraping-out cavities of body organs, e.g. bones; for invasive removal or destruction of calculus using mechanical vibrations; for removing obstructions in blood vessels, not otherwise provided for using mechanical vibrations, e.g. ultrasonic shock waves in direct contact with, or very close to, the obstruction or concrement the ultrasound transducer being inside patient's body at the distal end of the catheter
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B17/00—Surgical instruments, devices or methods
    • A61B17/22—Implements for squeezing-off ulcers or the like on inner organs of the body; Implements for scraping-out cavities of body organs, e.g. bones; for invasive removal or destruction of calculus using mechanical vibrations; for removing obstructions in blood vessels, not otherwise provided for
    • A61B2017/22051—Implements for squeezing-off ulcers or the like on inner organs of the body; Implements for scraping-out cavities of body organs, e.g. bones; for invasive removal or destruction of calculus using mechanical vibrations; for removing obstructions in blood vessels, not otherwise provided for with an inflatable part, e.g. balloon, for positioning, blocking, or immobilisation
    • A61B2017/22062—Implements for squeezing-off ulcers or the like on inner organs of the body; Implements for scraping-out cavities of body organs, e.g. bones; for invasive removal or destruction of calculus using mechanical vibrations; for removing obstructions in blood vessels, not otherwise provided for with an inflatable part, e.g. balloon, for positioning, blocking, or immobilisation to be filled with liquid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B17/00—Surgical instruments, devices or methods
    • A61B17/22—Implements for squeezing-off ulcers or the like on inner organs of the body; Implements for scraping-out cavities of body organs, e.g. bones; for invasive removal or destruction of calculus using mechanical vibrations; for removing obstructions in blood vessels, not otherwise provided for
    • A61B2017/22082—Implements for squeezing-off ulcers or the like on inner organs of the body; Implements for scraping-out cavities of body organs, e.g. bones; for invasive removal or destruction of calculus using mechanical vibrations; for removing obstructions in blood vessels, not otherwise provided for after introduction of a substance
    • A61B2017/22088—Implements for squeezing-off ulcers or the like on inner organs of the body; Implements for scraping-out cavities of body organs, e.g. bones; for invasive removal or destruction of calculus using mechanical vibrations; for removing obstructions in blood vessels, not otherwise provided for after introduction of a substance ultrasound absorbing, drug activated by ultrasound

Definitions

  • the present invention relates to a method and catheter for treating biological tissues with light activated drugs, and more particularly, to a method and catheter for treating biological tissues by delivering a light activated drug to a biological tissue and exposing the light activated drug to ultrasound energy.
  • Light activated drugs are inactive until exposed to light of particular wavelengths, however, upon exposure to light of the appropriate wavelength, light activated drugs can exhibit a cytotoxic effect on the tissues where they are localized. It has been postulated that the cytotoxic effect is a result of the formation of singlet oxygen on exposure to light.
  • Photodynamic therapy begins with the systemic administration of a selected light activated drug to a patient.
  • the drug disperses throughout the body and is taken up by most tissues within the body. After a period of time usually between 3 and 48 hours, the drug clears from most normal tissue and is retained to a greater degree in lipid rich regions such as the liver, kidney, tumor and atheroma.
  • a light source such as a fiber optic, is then directed to a targeted tissue site which includes the light activated drug. The tissues of the tissue site are then exposed to light from the light source in order to activate any light activated drugs within the tissue site. The activation of the light activated drug causes tissue death within the tissue site.
  • the concentration of the light activated drug within the targeted tissue site is limited by the quantity of light activated drug administered.
  • the concentration of the light activated drug within a tissue site can also be limited by the degree of selective uptake of the light activated drug into the tissue site. Specifically, if the targeted tissue site does not selectively uptake the light activated drug, the concentration of light activated drug within the tissue site can be too low for an effective treatment.
  • An additional problem associated with photodynamic therapy concerns depth of treatment. Light cannot penetrate deeply into opaque tissues. As a result, the depth that light penetrates most tissue sites is limited. This limited depth can prevent photodynamic therapy from being used to treat tissues which are located deeply in the interior of a tissue site.
  • An object for an embodiment of the invention is causing tissue death within a tissue site.
  • a further object for an embodiment of the present invention is using a catheter to locally deliver a light activated drug to a tissue site and delivering ultrasound energy from an ultrasound element on the catheter to activate the light activated drug.
  • An additional object for an embodiment of the present invention is including the light activated drug in an aqueous solution, locally delivering the aqueous solution to a tissue site and delivering ultrasound energy to the light activated drug within the tissue site to activate the light activated drug.
  • Yet a further object for an embodiment of the present invention is including the light activated drug in an emulsion, systemically delivering the emulsion, providing the light activated drug sufficient time to localize within a tissue site and delivering ultrasound energy to the light activated drug within the tissue site to activate the light activated drug.
  • a further object for an embodiment of the present invention is including the light activated drug in liposomes, systemically delivering the liposomes, providing the light activated drug sufficient time to localize within a tissue site and delivering ultrasound energy to the light activated drug within the tissue site to activate the light activated drug.
  • An additional object for an embodiment of the present invention is including the light activated drug in an aqueous solution, systemically delivering the aqueous solution, providing the light activated drug sufficient time to localize within a tissue site and delivering ultrasound energy to the light activated drug within the tissue site to activate the light activated drug.
  • Another object for an embodiment of the present invention is coupling a site directing molecule to a light activated drug, locally delivering the light activated drug to a tissue site and activating the light activated drug within the tissue site.
  • Yet another object for an embodiment of the invention is providing a catheter for locally delivering a media including a light activated drug to a tissue site.
  • the catheter including an ultrasound assembly configured to activate the light activated drug within the tissue site.
  • a further object for an embodiment of the invention is providing a catheter for delivering a media including a light activated drug to a tissue site.
  • the catheter including an ultrasound assembly for reducing exposure of the light activated drug to ultrasound energy until the light activated drug has been delivered from within the catheter.
  • a kit for causing tissue death within a tissue site includes a media with a light activated drug activatable upon exposure to a particular level of ultrasound energy.
  • the kit also includes a catheter with a lumen coupled with a media delivery port through which the light activated drug can be locally delivered to the tissue site.
  • the ultrasound transducer is configured to transmit the level of ultr.asound energy which activates the light activated drug with sufficient power that the ultrasound energy can penetrate the tissue site.
  • a method for causing tissue death in a subdermal tissue site includes providing a catheter for locally delivering a light activated drug to the subdermal tissue site, the catheter including an ultrasound transducer.
  • the method also includes locally delivering the light activated drug to the tissue site; producing ultrasound energy from the ultrasound transducer; and directing the ultrasound energy to the subdermal tissue site following penetration of the light activated drug into the subdermal tissue site to activate at least a portion of the light activated drug within the subdermal tissue site.
  • a method for activating a light activated drug includes providing a catheter with an ultrasound transducer.
  • the method also includes introducing the light activated drug into a patient's body where a subdermal tissue site absorbs at least a portion of the light activated drug; producing ultrasound energy; directing the ultrasound energy to the light activated containing subdermal tissue site including the light activated drug; and activating at least a portion of the light activated drug in the subdermal selected tissue site.
  • a method for releasing a therapeutic from a microbubble is also disclosed.
  • the method includes providing a microbubble with a light activated drug activatable upon exposure to ultrasound energy; and delivering ultrasound energy to the microbubble at a frequency and intensity which activates the light activated drug to cause a rupture of the microbubble.
  • a microbubble is also disclosed.
  • the microbubble includes a substrate defining a shell of the microbubble and having a thickness permitting hydraulic transport of the microbubble.
  • the microbubble also includes a light activated drug activatable upon exposure to ultrasound energy. Activation of the light activated drug causes a disruption in the shell sufficient to cause a rupture of the microbubble.
  • the microbubble further includes a therapeutic releasable from the microbubble upon rupture of the microbubble and yielding a therapeutic effect upon release from the microbubble.
  • Figure 1A is a side view of a catheter for locally delivering a media including a light activated drug to a tissue site.
  • Figure IB is an axial cross section of an ultrasound assembly for use with the catheter shown in Figure 1 A.
  • Figure 1C is a lateral cross section of an ultrasound assembly for use with the catheter shown in Figure 1 A.
  • Figure 2A is a side view of a catheter having an elongated body and an ultrasound assembly which is flush with the elongated body.
  • Figure 2B is an axial cross section of the ultrasound assembly illustrated in Figure
  • Figure 2C is a lateral cross section of the ultrasound assembly illustrated in Figure
  • Figure 3 A illustrates a catheter with a utility lumen and a second utility lumen.
  • Figure 3B is .an axial cross section of the ultrasound assembly illustrated in the catheter of Figure 3 A.
  • Figure 4A is a side view of a catheter including a plurality of ultrasound assemblies.
  • Figure 4B is a cross section of an ultrasound assembly included on a catheter with a plurality of utility lumens.
  • Figure 4C is a cross section of an ultrasound assembly included on a catheter with a plurality of utility lumens.
  • Figure 5 A is a side view of a catheter including a balloon.
  • Figure 5B is a cross section of a catheter with a balloon which include an ultrasound assembly.
  • Figure 6A is a side view of a catheter with a balloon positioned distally relative to an ultrasound assembly.
  • Figure 6B is a side view of a catheter with an ultrasound assembly positioned distally relative to a balloon.
  • Figure 6C is a cross section of a catheter with an ultrasound assembly positioned at the distal end of the catheter.
  • Figure 7A is a side view of a catheter with a media delivery port positioned between an ultrasound assembly and a balloon.
  • Figure 7B is a side view of a catheter with an ultrasound assembly positioned between a media delivery port and a balloon.
  • Figure 7C is a cross section of a catheter with an ultrasound assembly positioned at the distal end of the catheter.
  • Figure 8A is a side view of a catheter including a media delivery port and an ultrasound assembly positioned between first and second balloons.
  • Figure 8B is a side view of a catheter including a media delivery port and an ultrasound assembly positioned between first and second balloons.
  • Figure 8C is a cross section of a balloon included on a catheter having a first and second balloon.
  • Figure 9A illustrates an ultrasound assembly positioned adjacent to a tissue site and microbubbles delivered via a utility lumen.
  • Figure 9B illustrates an ultrasound assembly positioned adjacent to a tissue site and a media delivered via a media delivery port.
  • Figure 9C illustrates an ultrasound assembly positioned adjacent to a tissue site and a media delivered via a media delivery port while a guidewire is positioned in a utility lumen.
  • Figure 9D illustrates a catheter including a balloon positioned adjacent to a tissue site.
  • Figure 9E illustrates a catheter including a balloon expanded into contact with a tissue site.
  • Figure 9F illustrates a catheter with an ultrasound assembly outside a balloon positioned at a tissue site.
  • Figure 9G illustrates the balloon of Figure 9F expanded into contact with a vessel so as to occlude the vessel.
  • Figure 9H illustrates a catheter with an ultr.asound assembly outside a first and second balloon positioned at a tissue site.
  • Figure 91 illustrates the first and second balloon of Figure 9H expanded into contact with a vessel so as to occlude the vessel.
  • Figure 10A is a cross section of an ultrasound assembly according to the present invention.
  • Figure 1 OB is a cross section of an ultrasound assembly according to the present invention.
  • Figure IOC illustrates a support member with integral supports.
  • Figure 10D illustrates a support member which is supported by an outer coating.
  • Figure 11 A is a cross section of an ultrasound assembly including two concentric ultrasound transducers in contact with one another.
  • Figure 1 IB is a cross section of an ulfrasound assembly including two separated and concentric ultrasound transducers.
  • Figure 11C is a cross section of an ultrasound assembly including two ultrasound transducers where a chamber is defined between one of the ultrasound transducers and an elongated body.
  • Figure 1 ID is a cross section of an ulfrasound assembly including two longitudinally adjacent ultrasound transducers in physical contact with one another.
  • Figure 1 IE is a cross section of an ultrasound assembly including two separated and longitudinally adjacent ultrasound transducers.
  • Figure 1 IF is a cross section of an ultrasound assembly including two longitudinally adjacent ultrasound transducers with a single chamber positioned between both ultrasound transducers and an elongated body.
  • Figure 1 IG is a cross section of an ultrasound assembly including two longitudinally adjacent ultrasound transducers with different chambers positioned between each ultrasound transducers and an elongated body.
  • Figure 1 IH is a cross section of an ultrasound assembly including two longitudinally adjacent ultrasound transducers in contact with one another and having a single chamber positioned between each ultrasound transducers and an elongated body.
  • Figure 12A is a cross section of a catheter which includes an ultrasound assembly module which is independent of a first catheter component and a second catheter component.
  • Figure 12B illustrates the first and second catheter components coupled with the ultrasound assembly module.
  • Figure 12C is a cross section of an ultrasound assembly which is integral with a catheter.
  • Figure 13A is a cross section of an ultrasound assembly configured to radiate ultrasound energy in a radial direction.
  • the lines which drive the ultrasound transducer pass through a utility lumen in the catheter.
  • Figure 13B is a cross section of an ultrasound assembly configured to radiate ultrasound energy in a radial direction.
  • the lines which drive the ultrasound transducer pass through line lumens in the catheter.
  • Figure 13C is a cross section of an ultrasound assembly configured to longitudinally radiate ultrasound energy. The distal portion of one line travels proximally through the outer coating.
  • Figure 13D is a cross section of an ultrasound assembly configured to longitudinally transmit ultrasound energy. The distal portion of one line travels proximally through a line lumen in the catheter.
  • Figure 14A illustrates ultrasound transducers connected in parallel.
  • Figure 14B illustrates ultrasound transducers connected in series.
  • Figure 14C illustrates ultrasound transducers connected with a common line.
  • Figure 15 illustrates a circuit for electrically coupling temperature sensors.
  • Figure 17A illustrates pyrrole-based macrocyclic classes of light emitting drugs.
  • Figure 17B illustrates possible texaphyrin derivation sites.
  • Figure 18 illustrates the formula of preferred light emitting drugs for use with media including microbubbles.
  • Figure 19 illustrates a formula for a porphyrin group.
  • Figure 20 illustrates the formula of four preferred forms of the hydro- monobenzoporphyrin derivatives of the green porphyrins illustrated in formulae 3 and 4 of Figure 18.
  • Figure 21 illustrates the formulae for specific examples of pyrrole-based macrocycle derivatives and xanthene derivatives which are preferred for inclusion in microbubbles to enhance rupture of the microbubbles upon activation.
  • Figure 22 schematically summarizes the synthesis of an oligonucleotide conjugate of a texaphyrin metal complex.
  • Figure 23 illustrates the covalent coupling of texaphyrin metal complexes with amine, thiol, or hydroxy linked oligonucleotides.
  • Figure 24 illustrates the synthesis of diformyl monoic acid and oligonucleotide conjugate.
  • Figure 25 illustrates the synthesis of a texaphyrin based light activated drug.
  • Figure 26 illustrates the formula for tin ethyl etiopurpurin (SnEt 2 ).
  • the present invention relates to a method and catheter for delivering a light activated drug to a tissue site and delivering ultrasound energy to the light activated drug within the tissue site. Since many light activated drugs are also activated by ultrasound energy, the delivery of ultrasound energy to the light activated drug activates the light activated drug within the tissue site. Similar to activation of a light activated drug by light, activation by ultrasound causes death of tissues within the tissue site. The tissue death is believed to result from the release of a singlet oxygen.
  • Suitable tissue sites include, but are not limited to, atheroma, cancerous tumors, thrombi and potential restenosis sites.
  • a potential restenosis site is a tissue site where restenosis is likely to occur such as the portion of vessels previously treated by balloon angioplasty.
  • ultrasound energy can be transmitted through opaque tissues. As a result, the ultrasound energy can be used to treat tissues which are deeper within a tissue site than could be treated via light activation.
  • One explanation for the activation of light activated drugs via the application of ultrasound is a result of cavitation. Cavitation is known to occur when ultrasonic energy above a certain threshold is applied to a liquid.
  • Cavitation and Bubble Dynamics results when gas dissolved in a solution forms bubbles under certain types of acoustic vibration. Cavitation can also occur when small bubbles already present in the solution oscillate or repeatedly enlarge and contract to become bubbles. When the size of these cavitation bubbles reaches a size that cannot be maintained, they suddenly collapse and release various types of energy.
  • the various types of energy include, but are not limited to, mechanical energy, visible light, ultraviolet light and other types of electromagnetic radiation. Heat, plasma, magnetic fields, shock waves, free radicals, heat and other forms of energy are also thought to be generated locally.
  • the light activated drug is believed to be activated by at least one of the various forms of energy generated at the time of cavitation collapse.
  • the delivery of light activated drug to the tissue site can be through traditional systemic administration of a media including the light activated drug or can be performed through localized delivery of the media. Localized delivery can be achieved through injection into the tissue site or through other traditional localized delivery techniques.
  • a preferred delivery technique is using a catheter which includes a media delivery lumen coupled with a media delivery port. The catheter can be positioned such that the media delivery port is within the tissue site or is adjacent to the tissue site via traditional over- the-guidewire techniques. The media can then be locally delivered to the tissue site through the media delivery port.
  • the localized delivery of the light activated drug to the tissue sight serves to localize the light activated drug within the tissue site and can reduce the amount of light activated drug which concentrates in tissues outside the tissue site. Further, localized delivery of the light activated drug can serve to increase the concentration of the light activated drug within the tissue site above levels which would be achieved through systemic delivery of the light activated drug. Alternatively, the same concentration of light activated drug within the tissue site as would occur through systemic administration can be achieved by introducing smaller amounts of light activated drug into a patient's body.
  • Localized delivery of the light activated drug also permits treatment of tissue sites which do not have selective uptake of the light activated drug.
  • many light activated drugs such as the texaphyrins, are taken up by most tissues within the body and later localize within lipid rich tissues.
  • a non-lipid rich tissue site can be treated by delivering the ultrasound energy to the tissue site before the light activated drug has an opportunity to localize in lipid rich tissues.
  • Localized delivery is also advantageous when the tissue site is lipid rich such as in an atheroma or a tumor.
  • the localized delivery of the light activated drug combined with the inherent affinity of the light activated drug for tissue site can result in a high degree of localization of the light activated drug within lipid rich tissue sites.
  • the light activated drug can be coupled with a sight directing molecule to form a light activated drug conjugate.
  • the site directing molecule is chosen so the light activated drug conjugate specifically binds with the tissue site when the light activated drug conjugate is contacted with the tissue site under physiological conditions of temperature and pH.
  • the specific binding may result from specific electrostatic, hydrophobic, entropic, or other interactions between certain residues on the conjugate and specific residues on the tissue site.
  • the light activated drug includes an oligonucleotide acting as a site specific molecule coupled with a texaphyrin.
  • the oligonucleotide can have an affinity for a targeted site on a DNA strand.
  • the oligonucleotide can be designed to have complementary Watson-Crick base pairing with the targeted DNA site. Activation of the light activated drug after the conjugate has bound the targeted
  • the DNA site can cause cleavage of the DNA strand at the targeted DNA site.
  • the activated rug conjugate can be used for cleavage of targeted DNA sites.
  • the light activated conjugate can be targeted to a site on viral DNA where activation of the light activated conjugate causes the virus to be killed. Similarly, the light activated conjugate can be targeted to oncogenes.
  • targeted DNA cleavage include, but are not limited to, antisense applications, specific cleavage and subsequent recombination of DNA; destruction of viral DNA; construction of probes for controlling gene expression at the cellular level and for diagnosis; and cleavage of DNA in footprinting analyses, DNA sequencing, chromosome analysis, gene isolation, recombinant DNA manipulations, mapping of large genomes and chromosomes, in chemotherapy and in site directing mutagenesis.
  • the light activated drug includes a hormone.
  • the hormone may be targeted to a particular biological receptor which is localized at the tissue site.
  • the light activated drug can be included within several media suitable for delivery into the body. Many Ught activated drugs are known to have low water solubilities of less than 100 mg/L. As a result, achieving the desired concentration of light activated drug in an aqueous solution media for systemic delivery can often be difficult. However, localized delivery of the light activated drug requires a lower concentration of light activated drug within the media. As a result, when the light activated drug is delivered locally, the light activated drug can be included in an aqueous solution.
  • the media can also be an emulsion which includes a lipoid as a hydrophobic phase dispersed in a hydrophilic phase. These emulsions provide a media which is safe for delivery into the body with an effective concentration of light activated drug.
  • the media can also include microbubbles comprised from a substrate which forms a shell.
  • Suitable substrates for the microbubble include, but are not limited to, biocom- patible polymers, albumins, lipids, sugars or other substances.
  • the light activated drug can be enclosed within the microbubble, coupled with the shell and/or distributed in the media outside the microbubble.
  • a preferred microbubble comprises a lipid substrate such as liposome. Systemic admimstration of liposomes with light activated drug has been shown to result in an increased accumulation and more prolonged retention of light activated drugs within cultured malignant cells and within tumors in vivo. Jori et al., Br. J.
  • Including a light activated drug with the microbubbles has numerous advantages over microbubbles without light activated drug.
  • the microbubbles After administration of microbubbles to a patient, the microbubbles often must be ruptured to achieve their therapeutic effects.
  • One technique for rupturing microbubbles has been to expose the microbubbles to ultrasound energy.
  • ultrasound energy of undesirably high intensity is frequently required to break the microbubbles.
  • the ultrasound energy frequently must be matched to the resonant frequency of the microbubbles.
  • rupturing the microbubbles with ultr.asound can present numerous challenges.
  • Activating a light activated drug within the microbubble and/or in the substrate of the microbubble can cause the microbubble to rupture.
  • Activation of the light activated drug is believed to cause a disturbance which disrupts the shell of the microbubble enough to cause the microbubble to rupture. This disruption occurs when the light activated drug is coupled with the shell of the microbubble or is entirely within the microbubble. This disruption is also believed to occur when light activated drug located the media outside the microbubbles is activated in proximity of the microbubble.
  • microbubbles can be ruptured by activating light activated drugs and without matching the ultrasound frequency to the resonant frequency of the microbubble.
  • a more efficient ruptureding of microbubbles can be achieved by delivering a level of ultrasound energy which is appropriate to activate the light activated drug and which is matched to the resonant frequency of the microbubble.
  • the cavitation threshold can require an ultrasound intensity which is lower than the intensity required to rupture microbubbles without light activated drugs. As a result, including light activated drug with microbubbles can reduce the intensity of ultrasound energy required to rupture the microbubble.
  • the threshold value of cavitation is also reduced in the proximity of many light activated drugs.
  • the light activated drug encourages cavitation in the proximity of the light activated drug.
  • the interior of the microbubbles may include a gas or may be devoid of gas.
  • a gas When a gas is present, the gas can occupy any portion of the microbubble's volume but preferably occupies 0.01-50% of the volume of the microbubble interior, more preferably 5-30% and most preferably 10-20%.
  • the volume of gas is less than 0.01% of the volume, cavitation can be hindered and when the volume of gas is greater than 50% the structural integrity of the microbubble shell can become too weak for the microbubble to be transported to the tissue site.
  • Suitable gasses for the interior of the microbubbles include, but are not limited to, biocompatible gasses such as air, nitrogen, carbon dioxide, oxygen, argon, fluorine, xenon, neon, helium, or combinations thereof.
  • biocompatible gasses such as air, nitrogen, carbon dioxide, oxygen, argon, fluorine, xenon, neon, helium, or combinations thereof.
  • the presence of tiny bubbles is known to reduce the cavitation threshold.
  • the presence of an appropriately sized gas bubble in the microbubble can enhance cavitation in the proximity of the light activated drug.
  • the microbubbles are preferably 0.01-100 ⁇ m in diameter. This size microbubble reduces excretion of the microbubble outside the body and also reduces interference of the microbubble with the flow of fluids within the body of the patient. Further, the microbubbles preferably have a shell thickness of 0.001-50 ⁇ m, 0.01-5 ⁇ m and 0.1-0.5 ⁇ m. This thickness provides the shells with sufficient thickness that the microbubble can withstand enough of the forces within the vasculature of a patient to be transported through at least a portion of the patient's vasculature. Similarly, the thickness can permit the microbubbles to be transported through a lumen in an apparatus such as a catheter. However, this thickness is also sufficiently thin that alteration of the ultrasound activated substance upon activation is sufficient to disrupt the shell of the microbubble and cause the microbubble to rupture.
  • Activating the light activated drug to rapture microbubbles cm cause the light activated drug to be released from the microbubble so the light activated drag can penetrate the tissue near the site of rapture. Further exposure of the light activated drug to ultrasound can activate the light activated drag within the tissue and cause death of the tissue as described above.
  • the microbubble can include a therapeutic in addition to the light activated drag. Activation of the light activated drag can serve to rapture the microbubble and release the therapeutic from the microbubble. As a result, the therapeutic is released in proximity to a tissue site by rapturing the microbubble in proximity to the tissue site. This is advantageous when the therapeutic can be detrimental when administered systemically.
  • a therapeutic such as cisplatin is known to kill cancerous tissues but is also known to kill other tissues throughout the body.
  • systemic administration of cisplatin can be detrimental.
  • microbubbles can serve to protect tissues from the therapeutic agent until the therapeutic agent is released from the carrier. For instance, when the therapeutic is enclosed within the interior of the microbubble, contact between the therapeutic agent and tissues outside the carrier is reduced. As a result, the carrier increases protection of tissues outside the carrier are protected from the therapeutic agent until the microbubble is raptured .and the therapeutic released.
  • the therapeutics may be encapsulated in the microbubbles, included in the shell of the microbubbles or in the media outside the microbubbles.
  • therapeutic as used herein, it is meant an agent having beneficial effect on the patient.
  • therapeutics which can be included with the microbubbles include, but are not limited to, hormone products such as, vasopressin and oxytocin and their derivatives, glucagon and thyroid agents as iodine products and anti-thyroid agents; cardiovascular products as chelating agents and mercurial diuretics and cardiac glycosides; respiratory products as xanthine derivatives (theophylline & aminophylline); anti-infectives as aminoglycosides, antifungals (amphotericin), penicillin and cephalosporin antibiotics, antiviral agents as Zidovudine, Ribavirin, Amantadine, Vidarabine, and Acyclovir, anti-helmintics, antimalarials, and antituberculous drags; biologicals as immune serums, antitoxins and antivenins, rabies prophylaxis products, bacterial vaccines, viral vaccines, toxoids; antineoplastics asnitrosureas, nitrogen mustards, antimetabolites (fluor
  • thrombolytic agents such as urokinase
  • coagulants such as thrombin
  • antineoplastic agents such as platinum compounds (e.g., spirop latin, cisplatin, and carboplatin), methotrexate, adriamycin, taxol, mitomycin, ansamitocin, bleomycin, cytosine arabinoside, arabinosyl adsnine, mercaptopolylysme, vincristine, busulfan, chlorambucil, melphalan (e.g.,PAM, L-PAM or phenylalanine mustard), mercaptopurine, mitotane, procarbazine hydrochloride dactinomycin (actinomycin D), daunorabicinhydrochloride, doxorubicin hydrochloride, mitomycin, plicamycin (mithramycin), aminoglutethimide, estram
  • platinum compounds e.g.,
  • Zidovudine Ribavirin andvidarabine monohydrate (adenine arabinoside, ara-A); antianginals such asdiltiazem, nifedipine, verapamil, erythrityl tefrariitrate, isosorbidedinitrate, nitroglycerin (glyceryl trinitrate) and pentaerythritoltetranitrate; anticoagulants such as phenprocoumon, heparin; antibiotics such as dapsone, chloramphenicol, neomycin, cefaclor, cefadroxil, cephalexin, cephradine erythromycin, clindamycin, lincomycin, amoxicillin, ampicillin, bacampicillin, carbenicillin, dicloxacillin, cyclacillin, picloxacillin, hetacillin, methicillin, nafcillin, oxacillin, penicillin G, penicillin
  • the therapeutic is a monoclonal antibody, such as a monoclonal antibody capable of binding to melanoma antigen.
  • RNA nucleic acids
  • DNA RNA
  • RNA Ribonucleic acid
  • DNA DNA
  • Types of genetic material that may be used include, for example, genes carried on expression vectors such as plasmids, phagemids, cosmids, yeast artificial chromosomes (YACs), and defective or "helper" viruses, antigene nucleic acids, both single and double stranded RNA and DNA and analogs thereof, such asphosphorothioate and phosphorodithioate oligodeoxynucleotides. Additionally, the genetic material may be combined, for example, with proteins or other polymers.
  • Examples of genetic therapeutics that may be included in the microbubbles include DNA encoding at least a portion of an HLAgene, DNA encoding at least a portion of dysfrophin, DNA encoding at least a portion of CFTR, DNA encoding at least a portion of IL-2, DNA encoding at least a portion of TNF, an antisense oligonucleotide capable of binding the DNA encoding at least a portion of Ras.
  • DNA encoding certain proteins may be used in the treatment of many different types of diseases.
  • adenosine deaminase may be provided to treat ADA deficiency
  • tumor necrosis factor and or interleukin-2 may be provided to treat advanced cancers
  • HDL receptor may be provided to treat liver disease
  • thymidine kinase may be provided to treat ovarian cancer, brain tumors, or HIV infection
  • HLA-B7 may be provided to treat malignant melanoma
  • interleukin-2 may be provided to treat neuroblastoma, malignant melanoma, or kidney cancer
  • interleukin-4 may be provided to treat cancer
  • HIV env may be provided to treat HIV infection
  • antisense ras/p53 may be provided to treat lung cancer
  • Factor VIII may be provided to treat Hemophilia B. See, for example, Science 258, 744-746.
  • a single microbubble may contain more than one therapeutic or microbubbles containing different therapeutics may be co-administered.
  • a monoclonal antibody capable of binding to melanoma antigen and an oligonucleotide encoding at least a portion of IL-2 may be administered in a single microbubble.
  • the phrase "at least a portion of,” as used herein, means that the entire gene need not be represented by the oligonucleotide, so long as the portion of the gene represented provides an effective block to gene expression.
  • microbubbles including a therapeutic can be administered before, after, during or intermittently with the administration of microbubbles without a therapeutic.
  • microbubbles without a therapeutic and microbubbles including a coagulant such as thrombin can be administered to a patient having liver cancer.
  • Activating the light activated drug included with the microbubbles serves to rapture the microbubbles and release the light activated drug and thrombin from the microbubbles. Further activation of the light activated drag can cause tissue death and the thrombin can cause coagulation in and around the damaged tissues.
  • Prodrags may be included in the microbubbles, and are included within the ambit of the term therapeutic, as used herein.
  • Prodrags are well known in the art and include inactive drug precursors which, when exposed to high temperature, metabolizing enzymes, cavitation and/or pressure, in the presence of oxygen or otherwise, or when released from the microbubbles, will form active drugs.
  • Such prodrags can be activated via the application of ultrasound to the prodrag-containing microbubbles with the resultant cavitation, heating, pressure, and/or release from the microbubbles. Suitable prodrags will be apparent to those skilled in the art, and are described, for example, in Sinkula et al., J. Pharm. Sci.
  • Prodrags may comprise inactive forms of the active drugs wherein a chemical group is present on the prodrug which renders it inactive and/or confers solubility or some other property to the drug.
  • the prodrags are generally inactive, but once the chemical group has been cleaved from the prodrug, by heat, cavitation, pressure, and/or by enzymes in the surrounding environment or otherwise, the active drug is generated.
  • prodrags are well described in the .art, .and comprise a wide variety of drags bound to chemical groups through bonds such as esters to short, medium or long chain aliphatic carbonates, hemiesters of organic phosphate, pyrophosphate, sulfate, amides, amino acids, azo bonds, carbamate, phosphamide, glucosiduronate, N-acetylglucosamine and beta-glucoside.
  • bonds such as esters to short, medium or long chain aliphatic carbonates, hemiesters of organic phosphate, pyrophosphate, sulfate, amides, amino acids, azo bonds, carbamate, phosphamide, glucosiduronate, N-acetylglucosamine and beta-glucoside.
  • drugs with the parent molecule and the reversible modification or linkage are as follows: convallatoxin with ketals, hydantoin with alkyl esters, chlorphenesin with glycine or alanins esters, acetaminophen with caffeine complex, acetylsalicylic acid with THAM salt, acetylsalicylic acid with acetamidophenyl ester, naloxone with sulfateester, 15-methylprostaglandin F sub 2 with methyl ester, procaine with polyethylene glycol, erythromycin with alkyl esters, clindamycin with alkylesters or phosphate esters, tetracycline with betains salts, 7-acylaminocephalosporins with ring-substituted acyloxybenzyl esters, nandrolone with phenylproprionate decanoate esters, estradiol with enol
  • Prodrags may also be designed as reversible drag derivatives and utilized as modifiers to enhance drag transport to site-specific tissues.
  • parent molecules with reversible modifications or linkages to influence transport to a site specific tissue and for enhanced therapeutic effect include isocyanate with haloalkyl nitrosurea, testosterone with propionateester, methotrexate (S-S'-dichloromethotrexate) with dialkyl esters, cytosine arabinoside with 5'-acylate, nitrogen mustard (2,2'-dichloro-N-methyldiethylamine), nitrogen mustard with aminomethyltetracycline, nitrogen mustard with cholesterol or estradiol ordehydroepiandrosterone esters and nitrogen mustard with azobenzene.
  • a particular chemical group to modify a given drug may be selected to influence the partitioning of the drug into either the shell or the interior of the microbubbles.
  • the bond selected to link the chemical group to the drag may be selected to have the desired rate of metabolism, e.g., hydrolysis in the case of ester bonds in the presence of serum esterases after release from the microbubbles.
  • the particular chemical group may be selected to influence the biodistribution of the drug employed in the microbubbles, e.g., N,N-bis(2-chloroethyl)-phosphorodiamidicacid with cyclic phosphoramide for ovarian adenocarcinoma.
  • the prodrags employed within the microbubbles may be designed to contain reversible derivatives which are utilized as modifiers of duration of activity to provide, prolong or depot action effects.
  • nicotinic acid may be modified with dextran .and carboxymethlydextran esters, streptomycin with alginic acid salt, dihydrostreptomycin with pamoate salt, cytarabine (ara-C) with 5'-adamantoats ester, ara-adenosine (ara-A) with 5-palmirate and 5'-benzoate esters, amphotericin B with methyl esters, testosterone with 17- beta -alkyl esters, estradiol with formate ester, prostaglandin with 2-(4-imidazolyl) ethylamine salt, dopamine with amino acid amides, chloramphenicol with mono- and bis(trimethylsilyl) ethers, and cycloguanil with pamoate salt
  • a depot or reservoir of long-acting drag may be released in vivo from the prodrug bearing microbubbles.
  • compounds which are generally thermally labile may be utilized to create toxic free radical compounds.
  • Compounds with azolinkages, peroxides and disulfide linkages which decompose with high temperature are preferred.
  • azo, peroxide or disulfide bond containing compounds are activated by cavitation and/or increased heating caused by the interaction of ultra with the microbubbles to create cascades of free radicals from these prodrags entrapped therein.
  • Exemplary drags or compounds which may be used to create free radical products include azo containing compounds such as azobenzene ⁇ '-azobisisobutyronitrile, azodicarbonamide, azolitmin, azomycin, azosemide, azosulfamide, azoxybenzene, aztreonam, sudan III, sulfachrysoidine, sulfamidochrysoidine and sulfasalazine, compoimds containing* disulfide bonds such as sulbentine, thiamine disulfide, thiolutin, thiram, compounds containing peroxides such as hydrogen peroxide and benzoylperoxide, 2,2'-azobisisobutyronitrile, 2,2'-azobis(2-amidopropane) dihydrochloride, and
  • a microbubble having oxygen gas on its interior should create extensive free radicals with cavitation.
  • metal ions from the transition series especially manganese, iron and copper can increase the rate of formation of reactive oxygen intermediates from oxygen.
  • the formation of free radicals in vivo can be increased.
  • These metal ions may be incorporated into the microbubbles as freesalts, as complexes, e.g., with EDTA,
  • DTP A, DOTA or desferrioxamine or asoxides of the metal ions.
  • derivatized complexes of the metal ions may be bound to lipid head groups, or lipophilic complexes of the ions may be incorporated into a lipid bilayer, for example.
  • thermal stimulation e.g., cavitation
  • these metal ions then will increase the rate of formation of reactive oxygen intermediates.
  • radiosensitizers such as metronidazole .and misonidazole may be incorporated into the gas-filled liposomes to create free radicals on thermal stimulation.
  • an acylated chemical group may be bound to a drag via an ester linkage which would readily cleave in vivo by enzymatic action in serum.
  • the acylated prodrag can be included in the microbubble. When the microbubble is raptured, the prodrag will then be exposed to the serum. The ester linkage is then cleaved by esterases in the serum, thereby generating the drug.
  • ultrasound may be utilized not only to activate the light activated drag so as to burst the gas-filled liposome, but also to cause thermal effects which may increase the rate of the chemical cleavage and the release of the active drug from the prodrug.
  • the microbubbles may also be designed so that there is a symmetric or an asymmetric distribution of the therapeutic both inside and outside of the microbubble.
  • the particular chemical stracture of the therapeutics may be selected or modified to achieve desired solubility such that the therapeutic may either be encapsulated within the interior of the microbubble or couple with the shell of the microbubble.
  • the shell-bound therapeutic may bear one or more acyl chains such that, when the microbubble is popped or heated or ruptured via cavitation, the acylated therapeutic may then leave the surface and or the therapeutic may be cleaved from the acyl chains chemical group.
  • other therapeutics may be formulated with a hydrophobic group which is aromatic or sterol in stracture to incorporate into the surface of the microbubble.
  • the liposomes can be "fast breaking".
  • fast breaking liposomes the light activated drug-liposome combination is stable in vitro but, when admimstered in vivo, the light activated drag is rapidly released into the bloodstream where it can associate with serum lipoproteins.
  • the localized delivery of liposomes combined with the fast breaking nature of the liposomes can result in localization of the light activated drag and/or the therapeutic in the tissues near the catheter.
  • the fast breaking liposomes can prevent the liposomes from leaving the vicinity of the catheter intact and then concentrating in non-targeted tissues such as the liver. Delivery of ultrasound energy from the catheter can also serve to break apart the liposomes after they have been delivered from the catheter.
  • a catheter for locally delivering a media including a light activated drug includes an elongated body with at least one utility lumen extending through the elongated body.
  • the utility lumens can be used to deliver the media including the light activated drug locally to a tissue site and or to receive a guidewire so the catheter can be guided to the tissue site.
  • the ultrasound assembly can include an ultrasound transducer designed to transmit ultrasound energy which activates the light activated drug.
  • a support member can support the ultrasound transducer adjacent to an outer surface of the elongated body so as to define a chamber between the ultrasound transducer and the elongated body.
  • the chamber can be filled with a material which creates a low acoustic impedance to reduce the exposure of at least one utility lumen within the elongated body to ultrasound energy delivered from the ulfrasound transducer.
  • the chamber can be filled with a material which absorbs, reflects or prevents fransmission of ulfrasound energy through the chamber.
  • the chamber can be evacuated to reduce transmission of ulfrasound energy through the chamber. Reducing the exposure of at least one lumen to the ultrasound energy reduces exposure of media delivered through the at least one lumen to the ulfrasound energy. As a result, the effect of the ultrasound energy on the light activated drug is reduced until the light activated drag has been delivered out of the catheter. Further, ultrasound energy is known to rapture microbubbles. As a result, when the media includes microbubbles, the chamber reduces the opportunity for the ultrasound energy to rapture the microbubbles within the catheter.
  • the support member can have ends which extend beyond the ultrasound member.
  • the chamber can be positioned adjacent to the entire longitudinal length of the ultrasound transducer and can extend beyond the ends of the ultrasound transducer. This configuration maximizes the portion of the ultrasound transducer which is adjacent to the chamber. Increasing the portion of ultr.asound transducer adjacent to the chamber can reduce the amount of ultrasound energy transmitted to the utility lumens.
  • the ultrasound assembly can include an outer coating over the ulfrasound transducer. Temperature sensors can be positioned in the outer coating adjacent to ultrasound transducer. This position of the temperature sensors feedback regarding the temperature adjacent to the ulfrasound transducers where the thermal energy has a reduced opportunity to dissipate. As a result, the temperature sensors provide a measure of the temperature on the exterior surface of the transducer.
  • FIGS 1 A- IB illustrates a catheter 10 for delivering a media including a light activated drag to a tissue site.
  • the catheter 10 includes an ultrasound assembly 12 for delivering ultrasound energy to light activated drag within the tissue site.
  • the catheter 10 includes an elongated body 14 with a utility lumen 16 extending through the elongated body 14.
  • the utility lumen 16 can receive a guidewire (not shown) so the catheter 10 can be threaded along the guidewire.
  • the utility lumen 16 can also be used for the delivering media which include a light activated drug.
  • a fiber optic can also be positioned in the utility lumen 16 to provide a view of the tissue site or to provide light to the tissue site. As a result, the catheter can also be used as an endoscope.
  • the ultrasound assembly 12 can also include an outer coating 18. Suitable outer coatings 18 include, but are not limited to, polyimide, parylene and polyester. An ultrasound transducer 20 is positioned within the outer coating 18. Suitable ultrasound transducers 20 include, but are not limited to, PZT-4D, PZT-4, PZT-8 and cylindrically shaped piezoceramics. When the ultrasound transducer 20 has a cylindrical shape, the ultrasound transducer 20 can encircle the elongated body 14 as illustrated in Figure lC.
  • One or more temperature sensors 22 can be positioned in the outer coating 18.
  • the temperature sensors 22 can be positioned adjacent to the ultrasound fransducer 20 to provide feedback regarding the temperature adjacent to the ulfrasound transducer 20.
  • the temperature sensors can be in electrical communication with an electrical coupling 24.
  • the electrical coupling 24 can be coupled with a feedback control system (not shown) which adjusts the level of the ultrasound energy delivered from the ultrasound transducer 20 in response to the temperature at the temperature sensors 22.
  • the catheter 10 can include a perfusion lumen 25.
  • Ther perfusion lumen 25 allows fluid to flow foom outside the catheter into the utility lumen 16.
  • fluid flow which is obstructed by the ulfr.asound assembly can continue through the perfusion lumen 25 and the utility lumen.
  • the ulfrasound assembly 12 can be flush with the elongated body 14.
  • the ultrasound transducer 20 and the temperature sensors 22 can be positioned within the elongated body 14. This configuration of elongated body 14 and ulfrasound transducer 20 can eliminate the need for the outer coating 18 illustrated in
  • the catheter 10 can also include a media delivery port 26, a media inlet port 28 and a second utility lumen 16A.
  • the media inlet port 28 is designed to be coupled with a media source (not shown). Media can be transported from the media source and through the media delivery port 26 via the second utility lumen
  • a guidewire can be left within the utility lumen 16 while media is delivered via the second utility lumen 16 A.
  • Figure 4A illustrates a catheter 10 including a plurality of ultrasound assemblies 12.
  • Figures 4B-4C are cross sections of a catheter 10 with a second utility lumen 16A coupled with the media delivery ports 26.
  • the second utility lumen 16 A can also be coupled with the media inlet port 28 illustrated in Figure 4A.
  • the media inlet port 28 is designed to be coupled with a media source (not shown). Media can be transported from the media source and through the media delivery ports 26 via the second utility lumen 16 A.
  • the catheter 10 can include a balloon 30 as illustrated in Figure 5 A. The balloon
  • the membrane 30 can be constructed from an impermeable material or a permeable membrane or a selectively permeable membrane which allows certain media to flow through the membrane while preventing other media from flowing through the membrane.
  • Suitable membranous materials for the balloon 30 include, but are not limited to cellulose, cellulose acetate, polyvinylchloride, polyolefin, polyurethane and polysulfone.
  • the membrane pore sizes are preferably 5 A-2 ⁇ m, more preferably 50 A-900 A and most preferably 100 A- 300 A in diameter.
  • an ultrasound assembly 12 a first media delivery port 26A and a second media delivery port 26B can be positioned within the balloon 30.
  • the first and second media delivery ports 26A, 26B are coupled with a second utility lumen
  • the second .and third utility lumens 16 A, 16B can be coupled with the same media inlet port 28 or with independent media inlet ports 28.
  • different media can be delivered via the second and third media delivery ports 26 A, 26B.
  • a medication media can be delivered via the third utility lumen 16B and an expansion media can be delivered via the second utility lumen 16A.
  • the medication media can include drugs or other medicaments which can provide a therapeutic effect.
  • the expansion media can serve to expand the balloon 30 or wet the membrane comprising the balloon 30. Wetting the membrane comprising the balloon 30 can cause a minimally permeable membrane to become permeable.
  • the ultrasound assembly 12 can be positioned outside the balloon 30 as illustrated in Figures 6A-6C.
  • the balloon 30 is positioned distally of the ulfrasound assembly 12 and in Figure 6B the ultrasound assembly 12 is positioned distally of the balloon 30.
  • Figure 6C is a cross section a catheter 10 with an ultrasound assembly 12 positioned outside the balloon 30.
  • the catheter includes a second utility lumen 16A coupled with a first media delivery port 26A.
  • the second utility lumen 16A can be used to deliver an expansion media and or a medication media to the balloon 30.
  • the balloon 30 is constructed from a permeable membrane, the medication media and/or the expansion media can pass through the balloon 30.
  • the balloon 30 when the balloon 30 is constructed from a selectively permeable membrane, particular components of the medication media and or the expansion media can pass through the balloon 30. Pressure can be used to drive the media or components of the media across the balloon 30. Other means such as phoresis can also be used to drive the media or components of the media across the balloon 30.
  • the ultrasound assembly 12 may be positioned at the distal end of the catheter 10.
  • the second utility lumen 16A can be used to deliver an expansion media and or a medication media to the balloon 30.
  • the utility lumen 16 can be used to deliver a medication media as well as to guide the catheter 10 along a guidewire.
  • the catheter 10 can include a second media delivery port 26B positioned outside the balloon.
  • the ultrasound assembly 12 and the second media delivery port 26B are positioned distally relative to a balloon 30, however, the balloon 30 can be positioned distally relative to the ultrasound assembly 12 and the second media delivery port 26B.
  • the ultrasound assembly 12 is positioned distally of the second media delivery port 26B and in Figure 7B the second media delivery port 26B is positioned distally of the ultrasound assembly 12.
  • Figure 7C is a cross section of the catheter 10 illustrated in Figure 7 A.
  • the catheter 10 includes first and second media delivery ports 26A, 26B coupled with a second utility lumen 16A .and third utility lumen 16B.
  • the second and third utility lumens 16A, 16B can be coupled with independent media inlet ports 28 (not shown).
  • the second utility lumen 16 A can be used to deliver an expansion media and/or a medication media to the balloon 30 while the third utility lumen 16B can be used to deliver a medication media through the second media delivery port 26B.
  • the catheter 10 can include a first balloon 30A and a second balloon 30B.
  • the ultrasound assembly 12 can be positioned between the first and second balloons 30A, 30B.
  • a second media delivery port 26B can optionally be positioned between the first and second balloons 30A, 30B.
  • the second media delivery port 26B is positioned distally relative to the ultrasound assembly and in Figure 8B the ultrasound assembly is positioned distally relative to the second media delivery port 26B.
  • Figure 8C is a cross section of the first balloon 30A illustrated in Figure 8B.
  • the catheter includes a second, third and fourth utility lumens 16 A, 16B, 16C.
  • the second utiUty lumen 16A is coupled with a first media delivery port 26A within the first balloon.
  • the third utility lumen 16B is coupled with the second media delivery port 26B and the fourth utility lumen 16C is coupled with a third media delivery port 26C in the second balloon 30B (not shown).
  • the second and fourth utility lumens 16A, 16C can be used to deliver expansion media and/or medication media to the first and second balloon 30A, 30B.
  • the second and fourth utility lumens 16A, 16C can be coupled with the same media inlet port or with independent media inlet ports (not shown). When the second and fourth utility lumens are coupled with the same media inlet port, the pressure within the first .and second balloons 30A, 30B will be similar.
  • the third utility lumen 16B can be coupled with an independent media inlet port and can be used to deliver a medication media via the second media delivery port 26B.
  • Figures 9A-9I illustrate operation of various embodiments of catheters 10 for delivering ultrasound energy to a light activated drug within a tissue site.
  • Figures 9A-9I illustrate the tissue site 32 as an atheroma in a vessel 34, however, it is contemplated that the catheter 10 can be used with other tissue sites 32 such as a tumor and that the catheter 10 can be positioned within the vasculature of the tumor.
  • the catheter 10 is illustrated as being within a vessel 34.
  • the catheter 10 can be positioned within the vessel 34 by applying conventional over-the-guidewire techniques and can be verified by including radiopaque markers upon the catheter 10.
  • the catheter 10 is positioned so the ultrasound assembly 12 is adjacent to a tissue site 32 within a vessel 34.
  • the guidewire is removed from the utility lumen 16 .and media can be delivered via the utility lumen 16 as illustrated by the arrows 36.
  • the media includes microbubbles
  • the media is delivered to the tissue site 32 via the utility lumen 16 and ultrasound energy 40 is delivered from the ultrasound assembly 12.
  • Suitable periods for delivering the ultrasound energy include., but are not limited to, 1 minute to three hours, 2 minutes to one hour and 10 - 30 minutes.
  • Suitable intensities for the ultrasound energy include, but are not limited to, 0.1-
  • Suitable frequencies for the ultrasound energy include, but are not lmited to, 10 kHz- 100 MHz and 10 kHz- 50 MHz but is preferably 20 kHz- 10 MHz.
  • Suitable ultrasound energies also include, but are not limited to 0.02 to 10 w/cm 2 at a frequency of about 20 KHz to about 10 MHz and more preferably about 0.3 W/cm 2 at a frequency of about 1.3 MHz.
  • the ultrasound energy can be intermittently switched between a first and second frequency to increase the efficiency of microbubble rapture and to increase activation of the light activated drug.
  • the ultrasound energy can be switched between about 100 kHz and about 270 kHz in short pulses of approximately 0.001-10 seconds duration.
  • the ultrasound energy can be switched between first and second intensities.
  • the first and second frequencies can be provided by different ultrasound transducers.
  • the first and second intensities can be provided by different ultrasound transducers.
  • each transducer can simultaneously transmit ultrasound energy with different intensity and/or frequency.
  • the delivery of ultrasound energy 40 can be before, after, during or intermittently with the delivery of the microbubbles 38.
  • the microbubbles 38 can be "fast breaking" so they rapture upon exiting the utility lumen and being exposed to the vessel 34.
  • the ultrasound energy from the ultrasound assembly 12 can cause the microbubbles 38 within the delivered media to rapture.
  • the ultrasound assembly can be designed to reduce the exposure of media within the catheter 10 to the ultrasound energy from the ultrasound assembly 12.
  • the catheter 10 is so designed, the number of microbubbles 38 which rapture within the catheter is reduced and the number of microbubbles 38 which rupture outside the catheter is increased. Delivery of the ulfrasound energy before delivery of the light activated drag can enhance absorption of the light activated drug into the tissue site.
  • a pre-determined time after delivery of the light activated drug can provide the light activated drug time to penetrate the tissue site.
  • the pre-determined time can be of sufficient duration that at least a portion of the light activated drag penetrates into the tissue site.
  • the pre-determined time can also be of sufficient duration that the light activated drug localizes within the lipid rich tissue of the atheroma.
  • Sufficient time between delivery of the media and the ultrasound energy include but are not limited to, 1 minute to 48 hours, 1 minute to 3 hours, 1 to 15 minutes .and 1 to 2 minutes.
  • the ultr.asound energy from the ultiasound assembly 12 can activate the light activated dug within the tissue site 32 so as to cause tissue death within the tissue site 32.
  • ultrasound energy 40 is delivered from the ultrasound transducer 20 and a media is delivered through the media delivery port 26 as illustrated by the arrows 36.
  • the delivery of ultrasound energy 40 can be before, after, during or intermittently with the delivery of the media via the media delivery port 26.
  • the guidewire 104 can remain in the utility lumen 16 during the delivery of the media via the media delivery ports 26.
  • the ultrasound assembly can be designed to reduce the transmission of the ultrasound energy into the utility lumen. Because the fransmission of ultrasound energy 40 into the utility lumen 16 is reduced, the change in the frequency of the ultrasound transducer 20 which is due to the presence of the guidewire in the utility lumen 16 is also reduced.
  • a catheter 10 including a balloon 30 is positioned with the balloon adjacent to the tissue site 32.
  • the balloon 30 is expanded into contact with the tissue site 32.
  • the catheter 10 can include a perfusion lumen which permits a continuous flow of fluid from the vessel through the utility lumen during the partial or full obstruction of the vessel by the balloon.
  • a media can be delivered to the tissue site 32 via the balloon 30.
  • the media can serve to wet the membrane or can include a drag or other medicament which provides a therapeutic effect.
  • Ultrasound energy 40 can be delivered from the ultrasound assembly 12 before, after, during or intermittently with the delivery of the media.
  • the ulfrasound energy 40 can serve to drive the media across the membrane via phonophoresis or can enhance the therapeutic effect of the media.
  • a catheter 10 with an ultrasound assembly 12 outside a balloon 30 is positioned at the tissue site 32 so the ultrasound .assembly 12 is adjacent to the tissue site 32.
  • a fluid within the vessel flows past the balloon as indicated by the arrow 42.
  • the balloon 30 is expanded into contact with the vessel 34.
  • the balloon 30 can be constructed from an impermeable material so the vessel 34 is occluded. As a result, the fluid flow through the vessel 34 is reduced or stopped.
  • a medication media is delivered through the utility lumen 16 and ultrasound energy 40 is delivered from the ultrasound assembly 12.
  • the medication media can be delivered via the media delivery port 26.
  • a first medication media can be delivered via the media delivery port 26 while a second medication media can be delivered via the utility lumen 16 or while a guidewire is positioned within the utility lumen 16.
  • the ultrasound energy 40 can be delivered from the ultrasound assembly 12 before, after, during or intermittently with the delivery of the media.
  • the occlusion of the vessel 34 before the delivery of the media can serve to prevent the media from being swept from the tissue site 32 by the fluid flow.
  • the balloon 30 illustrated in Figures 9F-9G is positioned proximally relative to the ulfrasound assembly 12, the fluid flow through the vessel 34 can also be reduced by expanding a single balloon 30 which is positioned distally relative to the ultrasound assembly 12.
  • a catheter 10 including a first balloon 30A and a second balloon 30B is positioned at a tissue site 32 so the ultrasound assembly 12 is positioned adjacent to the tissue site 32.
  • a fluid within the vessel 34 flows past the balloon 30 as indicated by the arrow 42.
  • the first .and second balloons 30A, 30B are expanded into contact with the vessel 34.
  • the first and second balloons 30A, 30B can be constructed from an impermeable material so the vessel 34 is occluded proximally and distally of the ultrasound assembly 12. As a result, the fluid flow adjacent to the tissue site 32 is reduced or stopped.
  • a medication media is delivered through the media delivery port 26 and ulfrasound energy 40 is delivered from the ulfrasound assembly 12.
  • the ultrasound energy 40 can be delivered from the ulfrasound assembly 12 before, after, during or intermittently with the delivery of the media.
  • the occlusion of the vessel 34 before the delivery of the media can serve to prevent the media from being swept from the tissue site 32 by the fluid flow.
  • the media can be systemically delivered.
  • the catheter 10 is positioned adjacent to the tissue site before, after or during the systemic administration of the media.
  • the ulfrasound energy can be delivered after the microbubbles have had sufficient time to reach the desired tissue site in sufficient concentrations.
  • a level of ultrasound which ruptures the microbubbles is then delivered from the ultrasound assembly. After rupture of the microbubbles, the delivery of ultrasound energy can be stopped to provide the light activated drag or other therapeutic time to penetrate the tissue site.
  • the delivery of the ultrasound energy can also be continuous to maximize the number of microbubbles which are burst.
  • the behavior of the light activated drag within the patient must be taken into consideration.
  • many light drags such as the macrocycles, initially disperse throughout the body and where they are taken up by most tissues. After a period of time, usually between 3 and 48 hours, the drag clears from most normal tissue and is retained to a greater degree in lipid rich regions such as the liver, kidney, tumor and atheroma.
  • the tissue site is not a lipid rich region, the ultrasound energy should be delivered to the tissue site within 3 to 48 hours of systemically administering the media.
  • improved results can be achieved by waiting 3 to 48 hours after systemic administration of the media before delivering the ultrasound energy.
  • Figure 10A provides a cross section of an ultrasound assembly which reduces fransmission of ultrasound energy from the ultrasound fransducer into the catheter.
  • the ultrasound assembly 12 includes a support member 44.
  • Suitable support members 44 include, but are not limited to, polyimide, polyester .and nylon.
  • the support member 44 can be attached to the ultrasound transducer 20.
  • Suitable means for attaching the ultrasound transducer 20 to the support member 44 include, but are not limited to, adhesive bonding and thermal bonding.
  • the support member 44 supports the ultrasound member 44 at an external surface 46 of the elongated body 14 such that a chamber 48 is defined between the ultrasound transducer 20 and the external surface 46 of the elongated body 14.
  • the chamber 48 preferably has a height from .25-10 ⁇ m, more preferably from .50-5 ⁇ m and most preferably from .0-1.5 ⁇ m.
  • the support member 44 can be supported by supports 50 positioned at the ends 52 of the support member 44 as illustrated in Figure 10B.
  • the supports 50 can be integral with the support member 44 as illusfrated in Figure IOC.
  • the outer coating 18 can serve as the supports as illustrated in Figure 10D.
  • the ends 52 of the support member 44 can extend beyond the ends 54 of the ultrasound transducer 20.
  • the supports 50 can be positioned beyond the ends 54 of the ultrasound transducer 20.
  • the chamber 48 can extend along the longitudinal length 56 of the ultrasound transducer 20, maximizing the portion of the ultrasound transducer 20 which is adjacent to the chamber 48.
  • the chamber 48 can be filled with a medium which absorbs ultrasound energy or which prevents transmission of ultrasound energy.
  • Suitable gaseous media for filling the chamber 48 include, but are not limited to, helium, argon, air and nitrogen.
  • Suitable solid media for filling the chamber 48 include, but are not limited to, silicon and rabber.
  • the chamber 48 can also be evacuated. Suitable pressures for an evacuated chamber 48 include, but are not limited to, negative pressures to -760 mm Hg.
  • the ultrasound assembly can include a second ultrasound transducer 20A as illusfrated in Figures 11 A- 11 H.
  • a second ultrasound transducer 20A as illusfrated in Figures 11 A- 11 H.
  • one ultrasound transducer encircles the other and in Figures 11D-1 IH the ultrasound transducers are longitudinally adjacent to one another.
  • the ultrasound transducers 20, 20A can be in contact with one another as illustrated in Figures 11 A, 1 IE and 1 IH or separated from one another as illustrated in Figures 11B-1 ID, 1 IF and 1 IG.
  • a single chamber 54 can be defined between the ulfrasound transducers 20, 20A and the external surface 46 of the elongated body 14 as illustrated in Figures 11C, 1 IF and 1 IG or a different chamber can be defined between each of the ulfrasound transducers 20, 20A and the external surface 46.
  • the ulfrasound transducers 20, 20A in Figures 11A-11C are illustrated as having the same longitudinal length, the longitudinal length may be different.
  • the different temperature sensors can be positioned adjacent to different ulfrasound transducers 20, 20 A.
  • the temperature adjacent to different ultrasound transduers 20, 20A can be detected and the level of ultrasound energy produced by each ulfrasound transducer adjusted in repsonse to the detected temerature.
  • the transducers 20, 20A may be constructed from the same or different materials. Both transducers 20,
  • one transducer 20A may be configured to radiate ultrasound energy in the same direction. Further, one transducer may be configured to transmit ultrasound energy in a radial direction and the other in a longitudinal direction in order to increase the angular spectrum over which ulfrasound energy can be simultaneously transmitted.
  • the ultrasound transducers can be configured to fransmit ultrasound energy having the same or different characteristics. The transmission of ultrasound energy with different characteristics allows the same ultrasound assemblies to be used to perform different functions. For instance, one ultrasound transducer can transmit a frequency which is appropriate for activating a light activated drag while the second ultrasound transducer transmits a frequency appropriate for enhancing penetration of a therapeutic agent into the treatment site.
  • the transducers can be operated independently or simultaneously.
  • the ultrasound assembly When the transducers are operated simultaneously, the ultrasound assembly produces a waveform which is more complex than a single ultrasound transducer. More complex waveforms can provide advantages such as more efficient rapture of microbubbles. It is also contemplated that the ultrasound assembly can include three or more ulfrasound transducers arranged similar to the transducers illustrated in Figures 11 A- 11 H.
  • the ultrasound assembly 12 can be a separate module 58 as illusfrated in Figures 12A-12B.
  • the catheter 10 includes a first catheter component 60 a second catheter component 62 and an ultrasound assembly module 58.
  • the first and second catheter components 60, 62 include component ends 64 which are complementary to the ultrasound assembly module ends 66.
  • the component ends 64 can be coupled with the ultrasound assembly module ends 66 as illustrated in Figure 12B. Suitable means for coupling the component ends 64 and the ultrasound assembly module ends 66 include, but are not limited to, adhesive, mechanical and thermal methods.
  • the ultrasound assembly 12 can be integral with the catheter 10 as illustrated in Figure 12C.
  • the outer coating 18 can have a diameter which is larger than the diameter of the elongated body 14 as illusfrated in Figure 10A or can be flush with the external surface 46 of the elongated body 14 as illusfrated in Figures 12A-12C.
  • the ultrasound assembly 12 can be electrically coupled to produce radial vibrations of the ulfrasound fransducer 20 as illusfrated in Figures 13A-13B.
  • a first line 68 is coupled with an outer surface 70 of the ultrasound fransducer 20 while a second line
  • the first and second lines 68, 72 can pass proximally through the utility lumen 16 as illustrated in Figure 13 A. Alternatively, the first and second lines 68, 72 can pass proximally through line lumens 76 within toe catheter 10 as illustrated in Figure 13B.
  • Suitable lines for the ultrasound transducer 20 include, but are not limited to, copper, gold and aluminum.
  • Suitable frequencies for the ultrasound energy delivered by the ultrasound transducer 20 include, but are not limited to, 20 KHz to 2 MHz.
  • the ultrasound assembly 12 can be electrically coupled to produce longitudinal vibrations of the ultrasound transducer 20 as illustrated in Figures 13C-13D.
  • a first line 68 is coupled with a first end 78 of the ultrasound transducer 20 while a second line 72 is coupled with a second end 80 of the ultrasound transducer 20.
  • the distal portion 82 of the second line 72 can pass through the outer coating 18 as illustrated in Figure 13C.
  • the distal portion 82 of the second line 72 can pass through line lumens 76 in the catheter 10 as illustrated in Figure 13D.
  • the first and second lines 68, 72 can pass proximally through the utility lumen 16.
  • the catheter 10 can include a plurality of ultrasound .assemblies.
  • each ultrasound transducer 20 can each be individually powered.
  • the elongated body 14 includes N ultrasound transducers 20, the elongated body 14 must include 2N lines to individually power N ultrasound transducers 20.
  • each of the ultrasound transducers 20 can also be electrically coupled in serial or in parallel as illustrated in Figures 14A- 14B. These .arrangements permit maximum flexibility as they require only 2 lines.
  • Each of the ultrasound transducers 20 receive power simultaneously whether the ultrasound transducers 20 .are in series or in parallel. When the ultrasound transducers 20 .are in series, less current is required to produce the same power from each ultrasound transducer
  • the reduced current allows smaller lines to be used to provide power to the ultrasound transducers 20 and accordingly increases the flexibility of the elongated body 14.
  • an ultrasound transducer 20 can break down and the remaining ultrasound transducers 20 will continue to operate.
  • a common line 84 can provide power to each ultrasound transducer 20 while each ultrasound transducer 20 has its own return line 86.
  • a particular ultrasound transducer 20 can be individually activated by closing a switch 88 to complete a circuit between the common line 84 and the particular ultrasound transducer's 20 return line 86. Once a switch 88 corresponding to a particular ultrasound transducer 20 has been closed, the amount of power supplied to the ultrasound transducer 20 can be adjusted with the corresponding potentiometer 90. Accordingly, an catheter 10 with N ultrasound transducers 20 requires only N+l lines and still permits independent control of the ultrasound transducers 20. This reduced number of lines increases the flexibility of the catheter 10. To improve the flexibility of the catheter 10, the individual return lines 86 can have diameters which are smaller than the common line 84 diameter.
  • the diameter of the individual return lines 86 can be the square root of N times smaller than the diameter of the common line 84.
  • the ultrasound assembly 12 can include at least one temperature sensor 22.
  • Suitable temperature sensors 22 include, but are not limited to, thermistors, thermocouples, resistance temperature detectors (RTD)s, and fiber optic temperature sensors 22 which use thermalchromic liquid crystals.
  • Suitable temperature sensor geometries include, but are not limited to, a point, patch, stripe and a band encircling the ultrasound transducer 20.
  • the ulfrasound assembly 12 includes a plurality of temperature sensors 22, the temperature sensors 22 can be electrically connected as illustrated in Figure 15.
  • Each temperature sensor 22 can be coupled with a common line 84 and then include its own return line 86. Accordingly, N+l lines can be used to independently sense the temperature at the temperature sensors 22 when N temperature sensors 22 are employed.
  • a suitable common line 84 can be constructed from Constantine and suitable return lines
  • the 86 can be constructed from copper.
  • the temperature at a particular temperature sensor 22 can be determined by closing a switch 88 to complete a circuit between the thermocouple's return line 86 and the common line 84.
  • the temperature sensors 22 are thermocouples, the temperature can be calculated from the voltage in the circuit.
  • the individual return lines 86 can have diameters which are smaller than the common line 84 diameter.
  • Each temperature sensor 22 can also be independently electrically coupled. Employing N independently electrically coupled temperature sensors 22 requires 2N lines to pass the length of the catheter 10. The catheter 10 flexibility can also be improved by using fiber optic based temperature sensors 22. The flexibility can be improved because only N fiber optics need to be employed sense the temperature at N temperature sensors 22.
  • the catheter 10 can be coupled with a feedback control system as illustrated in Figure 16.
  • the temperature at each temperature sensor 22 is monitored and the output power of an energy source adjusted accordingly.
  • the physician can, if desired, override the closed or open loop system.
  • the feedback control system includes an energy source 92, power circuits 94 and a power calculation device 96 coupled with each ultrasound fransducer 20.
  • a temperature measurement device 98 is coupled with the temperature sensors 22 on the catheter 10.
  • a processing unit 100 is coupled with the power calculation device 96, the power circuits 94 and a user interface and display 102.
  • the temperature at each temperature sensor 22 is determined at the temperature measurement device 98.
  • the processing unit 100 receives a signals indicating the determined temperatures from the temperature measurement device 98.
  • the determined temperatures can then be displayed to the user at the user interface and display 102.
  • the processing unit 100 includes logic for generating a temperature control signal.
  • the temperature control signal is proportional to the difference between the measured temperature .and a desired temperature.
  • the desired temperature can be determined by the user.
  • the user can set the predetermined temperature at the user interface and display 102.
  • the temperature control signal is received by the power circuits 94.
  • the power circuits 94 adjust the power level of the energy supplied to the ultrasound transducers 20 from the energy source 92. For instance, when the temperature control signal is above a particular level, the power supplied to a particular ulfrasound transducer 20 is reduced in proportion to the magnitude of the temperature control signal. Similarly, when the temperature control signal is below a particular level, the power supplied to a particular ultrasound transducer 20 is increased in proportion to the magnitude of the temperature control signal.
  • the processing unit 100 monitors the temperature sensors 22 and produces another temperature control signal which is received by the power circuits 94.
  • the processing unit 100 can also include safety control logic.
  • the safety control logic detects when the temperature at a temperature sensor 22 has exceeded a safety threshold.
  • the processing unit 100 can then provide a temperature control signal which causes the power circuits 94 to stop the delivery of energy from the energy source 92 to the ultrasound transducers 20.
  • the processing unit 100 also receives a power signal from the power calculation device 96.
  • the power signal can be used to determine the power being received by each ulfrasound fransducer 20. The determined power can then be displayed to the user on the user interface and display 102.
  • the feedback control system can maintain the tissue adjacent to the ultrasound transducers 20 within a desired temperature range for a selected period of time.
  • the ulfrasound transducers 20 can be electrically connected so each ultrasound transducer 20 can generate an independent output. The output maintains a selected energy at each ultrasound transducer 20 for a selected length of time.
  • the processing unit 100 can be a digital or analog controller, or a computer with software. When the processing unit 100 is a computer it can include a CPU coupled through a system bus.
  • the user interface and display 102 can be a mouse, keyboard, a disk drive, or other non- volatile memory systems, a display monitor, and other peripherals, as are known in the art. Also coupled to the bus is a program memory and a data memory.
  • a profile of the power delivered to each ulfrasound transducer 20 can be incorporated in the processing unit 100 and a preset amount of energy to be delivered may also be profiled. The power delivered to each ultrasound transducer 20 can then be adjusted according to the profiles.
  • Suitable light activated drags include, but are not limited to, fluorescein, merocyanin.
  • preferred light activated drags include xanthene and its derivatives and the photoreactive pyrrole-derived macrocycles and their derivatives due to a reduced toxicity and an increased biological affinity.
  • Suitable photoreactive pyrrole- derived macrocycles include, but are not limited to, naturally occurring or synthetic porphyrins, naturally occurring or synthetic chlorins, naturally occurring or synthetic bacteriochlorins, synthetic isobateriochlorins, phthalocyanines, naphtalocyanines, and expanded pyrrole-based macrocyclic systems such as porphycenes, sapphyrins, and texaphyrins. Examples of suitable pyrrole-based macrocyclic classes are illustrated in
  • Figure 17B illusfrates a formula for the derivatives of texaphyrin: where M is H, CH 3 , a divalent metal cation selected from the group consisting of Ca(II), Mn(II), Co(II), Ni( ⁇ ), Zn(II),
  • CdiTI Hg(II), Fe(II), Sm(II), and UO(II) or a trivalent metal cation selected from the group consisting of Mn(III), Co(III), Ni(III), Fe(III), Ho(III), Ce( ⁇ i), Y(III), In(HI), Pr(UI), Nd(i ⁇ ), Sm(m), Eu(III), Gd(m), Tb(III), Dy(III), Er(III), Tm(III), Yb(IH), Lu(i ⁇ ), La(i ⁇ ), and U(III).
  • Preferred metals include Lu(III), Dy(III), Eu( ⁇ i), or Gd(i ⁇ ).
  • Rstrich R 2 , R 3 , R 4 , R 5 and R can independently be hydrogen, hydroxyl, alkyl, hydroxyalkyl, alkoxy, hydroxyalkoxy, saccharide, carboxyalkyl, carboxyamidealkyl, a site-directing molecule, or a linker to a site-directing molecule where at least one of R,, R 2 , R 3 , R 4 , R 5 and R ⁇ is hydroxyl, hydroxyalkoxy, saccharide, alkoxy, carboxyalkyl, carboxyamidealkyl, hydroxyalkyl, a site-directing molecule or a couple to a site-directing molecule; and N is an integer less than or equal to 2.
  • a preferred paramagnetic metal complex is the Gd(III) complex of 4,5-diethyl- 10,23 -dimethyl-9,24-bis(3-hydroxypropyl)- 16,17-bis[2-(2- methoxyethoxy)ethoxy]ethoxy-13,20,25,26,27- ⁇ entaazapentacyclo [20.2.1.1 3>6 .1 8 ' 11 .0 14 - 19 ] heptacosa-1,3,5,7,9,11 (27),12,14(19),15,17,20,22(25),23-tridecaene (“GdT2BET”) and a preferred diamagnetic metal complex is the Lu(III) complex of 4,5-diethyl- 10,23- dimethyl-9,24-bis(3-hydroxypropyl)-16,17-bis[2-[2-(2-methoxyethoxy)ethoxy]ethoxy]- 13,20,25,26,27-pentaazapentacyclo[20.2.
  • R t , R 2 , R 3 , R 4 , R 5 and R 6 may also independently be amino, carboxy, carboxamide, ester, amide sulfonato, aminoalkyl, sulfonatoalkyl, amidealkyl, aryl, etheramide or equivalent formulae conferring the desired properties.
  • at least one of R,, R 2 , R 3 , R 4 , R 5 and P ⁇ is a site-directing molecule or is a couple to a site- directing molecule.
  • R groups on the benzene ring portion of the molecule such as oligonucleotides, one skilled in the art would realize that derivatization at one position on the benzene potion is more preferred.
  • Hydroxyalkyl means alkyl groups having hydroxyl groups attached.
  • Alkoxy means alkyl groups attached to an oxygen.
  • Hydroxyalkoxy means alkyl groups having ether or ester linkages, hydroxyl groups, substituted hydroxyl groups, carboxyl groups, substituted carboxyl groups or the like.
  • Saccharide includes oxidized, reduced or substituted saccharide; hexoses such as D-glucose, D-mannose or D-galactose; pentoses such as D-ribose or D-arabinose; ketoses such as D-ribulose or D-fructose; disaccharides such as sucrose, lactose, or maltose; derivatives such as acetals, amines, and phosphorylated sugars; oligosacchrides, as well as open chain forms of various sugars, and the like.
  • amine-derivatized sugars are galactosamine, glucosamine, and sialic acid.
  • Carboxyamidealkyl means alkyl groups with secondary or tertiary amide linkages or the like.
  • Carboxyalkyl means alkyl groups having hydroxyl groups, carboxyl or amide substituted ethers, ester linkages, tertiary amide linkages removed from the ether or the like.
  • hydroxyalkoxy may be alkyl having independently hydroxy substituents and ether branches or may be C ( ⁇ . ⁇ ) H ((2 ⁇ +1) . 2 - ) O x O J , or
  • hydroxyalkoxy or saccharide may be C ⁇ H ((2 ⁇ + ⁇ ) .
  • n is a positive integer from 1 to 10
  • y is zero or a positive integer less than ((2n + 1) - q)
  • q is zero or a positive integer less than or equal to 2n+l
  • R ⁇ is independently H, alkyl, hydroxyalkyl, saccharide, C (m . w) H ((2m+1) - 2M , ) O w O-, 0 2 CC (m . VV ⁇ ⁇ D . 2W) 0 W 0 2 or N(R)OCC (m .
  • m is a positive integer from 1 to 10
  • w is zero or a positive integer less than or equal to m
  • z is zero or a positive integer less than or equal to ((2m+l) - 2w)
  • R is H, alkyl, hydroxyalkyl, or C m H ((2m+ i).
  • m is a positive integer from 1 to 10
  • z is zero or a positive integer less than ((2m+l) - r)
  • r is zero or a positive integer less than or equal to 2m+l
  • R* is independently H, alkyl, hydroxyalkyl, or saccharide.
  • Carboxyamidealkyl may be alkyl having secondary or tertiary amide linkages or (CH 2 ) ⁇ CONHR ⁇ , 0(CH 2 )-CONHR a , (CH 2 )-CON(R a )2, or 0(CH 2 ) n CON(R°)2 where n is a positive integer from 1 to 10, and R is independently H, alkyl, hydroxyalkyl, saccharide, O 2 CC (m . W) H ((2m+1) . 2lv) 0 w 0 z , N(R)OCC (m . H , ) H ((2m+1) . 2w) 0 H ,0-, or a site- directing molecule.
  • m is a positive integer from 1 to 10
  • w is zero or a positive integer less than or equal to ((2M+1) - 2w)
  • R is H, alkyl, hydroxyalkyl, or C m H ((2m+1) . r) 0-R''-.
  • m is a positive integer from 1 to 10
  • w is zero or a positive integer less than or equal to m
  • z is zero or a positive integer less than or equal to ((2M+1)
  • R ⁇ is an oligonucleotide.
  • Carboxyalkyl may be alkyl having a carboxyl substituted ether, an amide substituted ether or a tertiary amide removed from an ether or C-H ((2 _ +1) . ⁇ -) 0R c 1- or OC, ⁇ H ((2 ⁇ +1) . q) fi.
  • n is a positive integer from 1 to 10; y is zero or a positive integer less than ((2n+l) - q), q is zero or a positive integer less than or equal to 2n+l, and R c is (CH 2 ) administratC0 2 R'', (CH 2 ) ⁇ COHR d , (CH 2 ) choirCON(R ,) 2 or a site -directing molecule.
  • n is a positive integer from 1 to 10
  • R d is independently H, alkyl, hydroxyalkyl, saccharide, C (m .
  • m is a positive integer from 1 to 10
  • z is zero or a positive integer less than ((2m+l) - 2w)
  • R is H, alkyl, hydroxyalkyl, or C ⁇ ,, . r) OJH b r .
  • m is a positive integer from 1 to 10
  • z is zero or a positive integer less than ((2m+l)
  • R c is an oligonucleotide.
  • Exemplary texaphyrins are listed in Table 1.
  • A19 YCOCH,-linker- site-directing molecule Y NH, O
  • A22 CH,(CH,),OH CH,CH, CH,CH, ⁇ (CH,),C ⁇ -h ⁇ sta ⁇ une Preferred pyrrole-based macrocycles include, but are not limited to the hydro- monobenzoporphy ⁇ ns (the so-called “ft porphy ⁇ ne” or "Gp" compounds) disclosed in U.S. Patent Numbers 4,920,143 and 4,883,790 which are incorporated herein by reference. Typically, these compounds are poorly water-soluble (less than 1 mg/ml) or water-insoluble. Gp is preferably selected from the group consisting of those compounds having one of the formulae A-
  • R 1 and R 2 can be independently selected from the group consisting of carbalkoxy (2-6C), alkyl (1-6C) sulfonyl, aryl (6- IOC), sulfonyl, aryl (6- IOC), cyano, and — CONR 5 CO— wherein R 5 is aryl (6- IOC) or alkyl (1-6C).
  • each of R 1 and R 2 is carbalkoxy (2-6C).
  • R 3 can be independently carboxyalkyl (2-6C) or a salt, amide, ester or acylhydrazone thereof, or is alkyl (1-6C).
  • R 3 is — CH 2 CH 2 COOH or a salt, amide, ester or acylhydrazone thereof.
  • R 4 is — CHCH 2 , — CHOR 4 wherein R 4' is H or alkyl (1-6C), optionally substituted with a hydrophilic substituent; —CHO; — COOR 4 , CH(OR ' )CH 3 ; CH(OR 4 ) CH 2 OR 4 ; — CH(SR 4 )CH 3 ; — HNR 4 2 )CH 3 ; — CH(CN) CH 3 ; — CH(COOR 4' )CH 3 ; — CH(OOCR 4' )CH 3 ; — CH(halo)CH 3 ;
  • R 4 consists of 1-3 tetrapyrrole-type nuclei of the formula — L — P, wherein — L — is selected from the group consisting of
  • P is a second Gp, which is one of the formulae A-F ( Figure 18) but lacks R 4 , or another porphyrin group.
  • P is another porphy ⁇ n group
  • P preferably has the formula illustrated in Figure 19: wherein each R is independently H or lower alkyl (1-4C); two of the four bonds shown as unoccupied on adjacent rings are joined to R 3 ; one of the remaining bonds shown as unoccupied is joined to R 4 ; and the other is joined to L; with the proviso that, if R 4 is — CHCH 2 , both R 3 groups cannot be carbalkoxyethyl.
  • the preparation and use of such compounds is disclosed in U.S. Pat. Nos. 4,920,143 and 4,883,790, which are hereby incorporated by reference.
  • BPD's are light activated drags that are designated as benzoporphyrin derivatives ("BPD's").
  • BPD's are hydrolyzed forms, or partially hydrolyzed forms, of the rearranged products of formula A-C or formula A-D, where one or both of the protected carboxyl groups of R 3 are hydrolyzed.
  • BPD-MA is particularly preferred.
  • activating a light activated drug included in a microbubble can enhance rupture of the microbubble.
  • Preferred light activated drags for including m a microbubble to enhance rupture of the microbubble include Hematporphyrin, Rose Bengal, Eosm Y, Erythrocin, Rhodamine B, and PHOTOFRIN.
  • the formulae for these preferred light activated drugs are illustrated m Figure 21 where Rose Bengal, Eosm Y, Erythrocin and Rhodamine B are xanthene de ⁇ vanves.
  • the light activated drag can be coupled with a site directing molecule to form a light activated drug conjugate.
  • Suitable site-directing molecules include, but are not limited to: polydeoxyribonucleotides, oligodeoxyribonucleotides, polyribonucleotide analogs, oligoribonucleotide analogs; polyamides including peptides having an affinity for a biological receptor and proteins such as antibodies; steroids and steroid derivatives; hormones such as estradiol or histamine; hormone mimics such as morphine and further macrocycles such as sapphyrins and rubyrins.
  • nucleotide refers to both naturally occurring and synthetic nucleotides, poly- and oligonucleotides and to analogs and derivatives thereof such as methylphosphonates, phosphotriesters, phosphorothioates, and phosphoramidates and the like.
  • the oligonucleotide may be derivatized at the bases, the sugars, the end of the chains, or at the phosphate groups of the backbone to promote in vivo stability. Modifications of the phosphate groups are preferred in one embodiment since phosphate linkages are sensitive to nuclease activity. Preferred derivatives are the methylphosphonates, phosphotriesters, phosphorothioates, and phosphoramidates. Additionally, the phosphate linkages may be completely substituted with non-phosphate linkages such as amide linkages. Appendages to the ends of the oligonucleotide chains also provide exonuclease resistance.
  • Sugar modifications may include alkyl groups attached to an oxygen of a ribose moiety in a ribonucleotide.
  • the alkyl group is preferably a methyl group and the methyl group is attached to the 2' oxygen of the ribose.
  • Other alkyl groups may be ethyl or propyl.
  • a linker may be used to couple the light activated drag with the site directing molecule.
  • exemplary linkers include, but are not limited to, amides, amine, thioether, ether, or phosphate covalent bonds as described in the examples for attachment of oligonucleotides.
  • an oligonucleotide or other site-directing molecules is covalently bonded to a texaphyrin or other light activated drags via a carbon- nitrogen, carbon-sulfur, or a carbon-oxygen bond.
  • the media can be an emulsion which includes a light activated drag.
  • the emulsions described below are suitable for delivery into a body since they avoid pharmaceutically undesirable organic solvents, solubilizers, oils or emulsifiers.
  • a wide range of light activated drag concentrations can be used in the emulsion. Suitable concentrations of light activated drag within the emulsion include, but are not limited to, approximately 0.01 to 1 gram 100 ml, preferably about 0.05 to about 0.5 gram/100 ml, and approximately 0.1 g/100 ml.
  • the emulsion includes a lipoid as a hydrophobic component dispersed in a hydrophilic phase.
  • the hydrophobic component of the emulsion comprises a pharmaceutically acceptable triglyceride, such as an oil or fat of a vegetable or animal nature, and preferably is selected from the group consisting of soybean oil, safflower oil, marine oil, black current seed oil, borage oil, palm kernel oil, cotton seed oil, corn oil, sunflower seed oil, olive oil or coconut oil. Physical mixtures of oils .and/or interesterfied mixtures can be employed.
  • the preferred oils are medium chain length triglycerides having C 8 — C 10 chain length and more preferably saturated.
  • the preferred triglyceride is a distillate obtained from coconut oil.
  • the hydrophobic content of the emulsion is preferably approximately 5 to 50 g/100 ml, more preferably about 10 to about 30 g/100 ml and approximately 20 g/100 ml of the emulsion.
  • the emulsion can also contains a stabilizer such as phosphatides, soybean phospholipids, nonionic block copolymers of polyoxethylene and polyoxpropylene (e.g. poloxamers), synthetic or semi-synthetic phospholipids, and the like.
  • the preferred stabilizer is purified egg yolk phospholipid.
  • the stabilizer is usually present in the composition in amounts of about 0.1 to about 10, and preferably about 0.3 to about 3 grams/100 ml, a typical example being about 1.5 grams/100 ml.
  • the emulsion can also include one or more bile acids salts as a costablizer.
  • the salts are pharmacologically acceptable salts of bile acids selected from the group of cholic acid, deoxycholic acid and gylcocholic acid, and preferably of cholic acid.
  • the salts are typically alkaline metal or alkaline earth metal salts and preferably sodium, potassium, calcium or magnesium salts, and most preferably, sodium salts. Mixtures of bile acid salts can be employed if desired.
  • the amount of bile acid salt employed is usually about 0.01 to about 1.0 and preferably about 0.05 to about 0.4 grams/100 ml, a typical example being about 0.2 grams/100 ml.
  • Suitable pH for the emulsion includes, but is not limited to approximately 7.5 to
  • the pH can be adjusted to the desired value, if necessary, by adding a pharmaceutically acceptable base, such as sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide and ammonium hydroxide.
  • a pharmaceutically acceptable base such as sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide and ammonium hydroxide.
  • Water can be added to the emulsion to achieve the desired concentration of various components within the emulsion.
  • the emulsion can include auxiliary ingredients for regulating the osmotic pressure to make the emulsion isotonic with the blood. Suitable auxiliary ingredients include, but are not limited to, auxiliary surfactants, isotonic agents, antioxidants, nutritive agents, trace elements and vitamins.
  • Suitable isotonic agents include, but are not limited to, glycerin, amino acids, such as alanine, histidine, glycine, .and/or sugar alcohols, such as xylitol, sorbitol and/or mannitol.
  • Suitable concenfrations for isotonic agents within the emulsion include, but are not limited to, approximately 0.2 to about 8.0 grams/100 ml and preferably about 0.4 to about 4 grams/100 ml and most preferably 1.5 to 2.5 gram/100 ml.
  • Antioxidants can be used to enhance the stability of the emulsion, a typical example being ⁇ -tocopherol.
  • Suitable concentrations for the antioxidants include, but are not limited to approximately 0.005 to 0.5 grams/100 ml, approximately 0.02 to about 0.2 grams/100 ml and most preferably approximately 0.05 to 0.15 grams/100 ml.
  • the emulsions can also contain auxiliary solvents, such as an alcohol, such as ethyl alcohol or benzyl alcohol, with ethyl alcohol being preferred.
  • auxiliary solvents such as an alcohol, such as ethyl alcohol or benzyl alcohol, with ethyl alcohol being preferred.
  • alcohol such as ethyl alcohol or benzyl alcohol
  • ethyl alcohol being preferred.
  • ethyl alcohol is typically present in amounts of about 0.1 to about 4.0, and preferably about 0.2 to about
  • the ethanol is advantageous since it facilitates dissolution of poorly water-soluble light activated drugs and especially those that form crystals which may be very difficult to dissolve in the hydrophobic phase. Accordingly, the ethanol must be added directly to the hydrophobic phase during preparation to be effective. For maximum effectiveness, the ethanol should constitute about 5% to 15% by weight of the hydrophobic phase. In particular, if ethanol constitutes less than 5% by weight of the hydrophobic phase, dissolution of the light activated drug can become unacceptably slow. When the ethanol concentration exceeds 15%, large (>5 ⁇ m diameter) and poorly emulsified oil droplets can form in the emulsion.
  • the particles in the emulsion are preferably less than about 5.0 ⁇ m in diameter, more preferably less than 2.0 ⁇ m in diameter and most preferably less than 0.5 ⁇ m or below.
  • a typical emulsion is prepared using the following technique.
  • the triglyceride oil is heated to 50°-70° C while sparging with nitrogen gas.
  • the required amounts of stabilizer (e.g. egg yolk phospholipids), bile acid salt, alcohol (e.g. ethanol), antioxidant (e.g. ⁇ -to-copherol) and light activated drag are added to the triglyceride while processing for about 5 to about 20 minutes with a high speed blender or overhead mixer to ensure complete dissolution or uniform suspension.
  • the required amounts of water and isotonic agent e.g. - glycerin
  • the aqueous phase is transferred into the prepared hydrophobic phase and high speed blending is continued for another 5 to 10 minutes to produce a uniform but coarse preemulsion (or premix).
  • This premix is then transferred to a conventional high pressure homogenizer (APV Gaulin) for emulsification at about 8,000-10,000 psi.
  • AAV Gaulin conventional high pressure homogenizer
  • the diameter of the dispersed oil droplets in the finished emulsion will be less than 5 ⁇ m, with a large proportion less than 1 ⁇ m.
  • the mean diameter of these oil droplets will be less than 1 ⁇ m, preferably from 0.2 to 0.5 ⁇ m.
  • the emulsion product is then filled into borosilicate (Type 1) glass vials which are stoppered, capped and terminally heat sterilized in a rotating steam autoclave at about 121° C.
  • the vehicle composition employed is chemically inert with respect to the incorporated pharmacologically active light activated drug.
  • the emulsions can exhibit very low toxicity following intravenous admimstration and exhibit no venous irritation and no pain on injection.
  • the emulsions exhibit mimmal physical and chemical changes (e.e. formation of non-emulsified surface oil) during controlled shake-testing on a horizontal platform.
  • the oil-in-water emulsions promote desirable pharmacoldnetics and tissue distribution of the light activated drug in vivo.
  • the light activated drag can also be delivered to the body in a media which includes microbubbles.
  • Suitable substrates for the microbubble include, but are not limited to, biocompatible polymers, albumins, lipids, sugars or other substances.
  • U.S. patent numbers 5,665,383 and 5,665, 382 teaches a method for synthesizing microbubbles with a polymeric substrate and is inco ⁇ orated herein by reference.
  • U.S. patent numbers 5,626,833 and 5,798,091 teach methods for synthesizing microbubbles with a surfactant substrate and are incorporated herein by reference.
  • a preferred microbubble has a lipid substrate.
  • U.S. patent numbers 5,772,929 teaches methods for synthesizing microbubbles with a lipid substrate.
  • U.S. patent numbers 5,776,429, 5,715 ,824 and 5 ,770,222 teach preferred methods for synthesizing microbubbles with a lipid substrate and a gas interior and are incorporated herein by reference.
  • Suitable microbubbles with a lipid substrate can be liposomes.
  • the liposomes can be unilamellar vesicles having a single membrane bilayer or multilamellar vesicles having multiple membrane bilayers, each bilayer being separated from the next by an aqueous layer.
  • a liposome bilayer is composed of two lipid monolayers having a hydrophobic
  • the formula of the membrane bilayer is such that the hydrophobic (nonpolar) "tails” of the lipid monolayers orient themselves towards the center of the bilayer, while the hydrophilic "heads” orient themselves toward the aqueous phase.
  • Either unilamellar or multilamellar or other types of liposomes may be used.
  • a hydrophilic light activated drag can be entrapped in the aqueous phase of the liposome before the drag is delivered into the patient. Alternatively, if the light activated drag is lipophilic, it may associate with the lipid bilayer. Liposomes may be used to help "target" the light activated drag to an active site or to solubilize hydrophobic light activated drugs. Light activated drags are typically hydrophobic and form stable drag- lipid complexes.
  • light activated drugs have low solubility in water at physiological pH's, but are also insoluble in (1) pharmaceutically acceptable aqueous- organic co-solvents, (2) aqueous polymeric solutions and (3) surfactant/micellar solutions.
  • such light activated drags can still be "solubilized” in a form suitable for delivery into a body by using a liposome composition.
  • a light activated drug BPD-MA See Formula A of Figure 20
  • BPD-MA can be "solubilized” at a concentration of about 2.0 mg/ml in aqueous solution using an appropriate mixture of phospholipids to form encapsulating liposomes.
  • the light activated drag can be included in many different types of liposomes, the following description discloses particular liposome compositions and methods for making the liposomes which are known to be "fast breaking".
  • fast breaking liposomes the light activated drug-liposome combination is stable in vitro but, when administered in vivo, the light activated drag is rapidly released into the bloodstream where it can associate with serum lipoproteins.
  • the localized delivery of liposomes combined with the fast breaking nature of the liposomes can result in localization of the light activated drag in the tissues near the catheter.
  • the fast breaking liposomes can prevent the liposomes from leaving the vicinity of the catheter intact and then concentrating in non-targeted tissues such as the liver.
  • Liposomes are typically formed spontaneously by adding water to a dry lipid film.
  • Liposomes which include light activated drags can include a mixture of the commonly encountered lipids dimyristoyl phosphatidyl choline (“DMPC”) and egg phosphatidyl glycerol (“EPG").
  • DMPC dimyristoyl phosphatidyl choline
  • EPG egg phosphatidyl glycerol
  • the presence of DMPC is important because DMPC is the major component in the composition to form liposomes which can solubilize and encapsulate insoluble light activated drags into a lipid bilayer.
  • EPG egg phosphatidyl glycerol
  • phospholipids in addition to DMPC and EPG, may also be present.
  • additional phospholipids that may also be incorporated into the liposomes include phosphatidyl cho lines (PCS), including mixtures of dipalmitoyl phosphatidyl choline (DPPC) and distearoyl phosphatidyl choline (DSPC).
  • PCS phosphatidyl cho lines
  • DPPC dipalmitoyl phosphatidyl choline
  • DSPC distearoyl phosphatidyl choline
  • suitable phosphatidyl glycerols include dimyristoyl phosphatidyl glycerol (DMPG), DLPG and the like.
  • DMPG dimyristoyl phosphatidyl glycerol
  • DLPG DLPG
  • Other types of suitable lipids that may be included are phosphatidyl ethanolamines
  • the molar ratio of the light activated drag to the DMPC/EPG phospho lipid mixture can be as low as 1 :7.0 or may contain a higher proportion of phospholipid, such as 1 : 7.5. Preferably, this molar ratio is 1 : 8 or more phospholipid, such as 1:10, 1:15, or 1 :20. This molar ratio depends upon the exact light activated drug being used, but will assure the presence of a sufficient number of DMPC and EPG lipid molecules to form a stable complex with many light activated drags.
  • the lipophilic phase of the lipid bilayer becomes saturated with light activated drag molecules. Then, any slight change in the process conditions can force some of the previously encapsulated light activated drug to leak out of the vesicle, onto the surface of the lipid bilayer, or even out into the aqueous phase.
  • the concentration of light activated drug is high enough, it can actually precipitate out from the aqueous layer and promote aggregation of the liposomes.
  • the more unencapsulated light activated drug that is present the higher the degree of aggregation.
  • the more aggregation the larger the mean particle size will be, and the more difficult aseptic or sterile filtration will be.
  • small changes in the molar ratio can be important in achieving proper filterability of the liposome composition.
  • lipid content can increase significantly the filterability of the liposome composition by increasing the ability to form and maintain small particles.
  • This is particularly advantageous when working with significant volumes of 500 ml, a liter, five liters, 40 liters, or more, as opposed to smaller batches of about 100-500 ml or less.
  • This volume effect is thought to occur because larger homogenizing devices tend to provide less efficient agitation than can be accomplished easily on a small scale. For example, a large size MicrofluidizerTM has a less efficient interaction chamber than that one of a smaller size.
  • a molar ratio of 1.05:3:5 BPD-MA:EPG:DMPC may provide marginally acceptable filterability in small batches of up to 500 ml.
  • a higher molar ratio of phospholipid provides more assurance of reliable aseptic filterability.
  • the substantial potency losses that are common in scale-up batches, due at least in part to filterability problems, can thus be avoided.
  • cryoprotective agent known to be useful in the art of preparing freeze-dried formulations, such as di- or polysaccharides or other bulking agents such as lysine, may be used.
  • isotonic agents typically added to maintain isomolarity with body fluids may be used.
  • a di-saccharide or polysaccharide is used and functions both as a cryoprotective agent and as an isotonic agent.
  • the particular combination of the phospholipids, DMPC and EPG, and a disaccharide or polysaccharide form a liposomal composition having liposomes of a particularly narrow particle size distribution.
  • a disaccharide or polysaccharide provides instantaneous hydration and the large surface area for depositing a thin film of the drag-phospholipid complex. This thin film provides for faster hydration so that, when the liposome is initially formed by adding the aqueous phase, the liposomes formed are of a smaller and more uniform particle size. This provides significant advantages in terms of manufacturing ease.
  • a saccharide when present in the composition, it is added after dry lipid film formation, as a part of the aqueous solution used in hydration.
  • a saccharide is added to the dry lipid film during hydration.
  • Disaccharides or polysaccharides are preferred to monosaccharides for this purpose.
  • no more than 4-5% monosaccharides could be added.
  • about 9-10% of a disaccharide can be used without generating an unacceptable osmotic pressure.
  • the higher amount of disaccharide provides for a larger surface area, which results in smaller particle sizes being formed during hydration of the lipid film.
  • the preferred liposomal composition comprises a disaccharide or polysaccharide, in addition to the light activated drug and the mixture of DMPC and EPG phospholipids.
  • the disaccharide or polysaccharide is preferably chosen from among the group consisting of lactose, frehalose, maltose, maltotriose, palatinose, lactulose or sucrose, with lactose or frehalose being preferred.
  • the liposomes comprise lactose or frehalose.
  • the disaccharide or polysaccharide is formulated in a preferred ratio of about 10-20 saccharide to 0.5-6.0 DMPC/EPG phospholipid mixture, respectively, even more preferably at a ratio from about 10 to 1.5-4.0.
  • a preferred but not limiting formulation is lactose or frehalose and a mixture of DMPC and EPG in a concentration ratio of about 10 to 0.94-1.88 to about 0.65-1.30, respectively.
  • liposomes having extremely small and narrow particle size ranges not only tends to yield liposomes having extremely small and narrow particle size ranges, but also provides a liposome composition in which light activated drugs, in a particular, may be stably incorporated in an efficient manner, i.e., with an encapsulation efficiency approaching 80-100%.
  • liposomes made with a saccharide typically exhibit improved physical and chemical stability, such that they can retain an incorporated light activated drag without leakage upon prolonged storage, either as a reconstituted liposomal or as a cryodesiccated powder.
  • antioxidants e.g., butylated hydroxytoluene, alphatocopherol and ascorbyl palmitate
  • pH buggering agents e.g., phosphates, glycine, and the like.
  • Liposomes containing a light activated drug may be prepared by combining the light activated drag and the DMPC and EPG phospholipids (and any other optional phospholipids or excipients, such as antioxidants) in the presence of an organic solvent.
  • Suitable organic solvents include any volatile organic solvent, such as diethyl ether, acetone, methylene chloride, chloroform, piperidine, piperidine-water mixtures, methanol, tert-butanol, dimethyl sulfoxide, N-methyl-2-pyrrolidone, and mixtures thereof.
  • the organic solvent is water-immiscible, such as methylene chloride, but water immiscibility is not required.
  • the solvent chosen should not only be able to dissolve all of the components of the lipid film, but should also not react with, or otherwise deleteriously affect, these components to any significant degree.
  • the organic solvent is then removed from the resulting solution to form a dry lipid film by any known laboratory technique that is not deleterious to the dry lipid film and the light activated drug.
  • the solvent is removed by placing the solution under a vacuum until the organic solvent is evaporated.
  • the solid residue is the dry lipid film.
  • the thickness of the lipid film is not critical, but usually varies from about 30 to about 45 mg/cm 2 , depending upon the amount of solid residual and the total area of the glass wall of the flask.
  • the film may be stored for an extended period of time, preferably not more than 4 to 21 days, prior to hydration. While the temperature during a lipid film storage period is also not an important factor, it is preferably below room temperature, most preferably in the range from about -20 to about 4° C.
  • the dry lipid film is then dispersed in an aqueous solution, preferably containing a disaccharide or polysaccharide, and homogenized to form the desired particle size.
  • aqueous solutions used during the hydration step include sterile water; a calcium- and magnesium- free, phosphate-buffered (pH 7.2-7.4) sodium chloride solution; a 9.75% w/v lactose solution; a lactose-saline solution; 5% dextrose solution; or any other physiologically acceptable aqueous solution of one or more electrolytes.
  • the aqueous solution is sterile.
  • the volume of aqueous solution used during hydration can vary greatly, but should not be so great as about 98% nor so small as about 30-40%. A typical range of useful volumes would be from about 75% to about 95%, preferably about 85% to about 90%.
  • the "therapeutically effective amount” can vary widely, depending on the tissue to be treated and whether it is coupled to a target-specific ligand, such as an antibody or an immunologically active fragment. It should be noted that the various parameters used for selective photodynamic therapy are interrelated. Therefore, the therapeutically effective amount should also be adjusted with respect to other parameters, for example, fluence, irradiance, duration of the light used in photodynamic therapy, and the time interval between administration of the light activated drag and the therapeutic irradiation. Generally, all of these parameters are adjusted to produce significant damage to tissue deemed undesirable, such as neovascular or tumor tissue, without significant damage to the surrounding tissue, or to enable the observation of such undesirable tissue without significant damage to the surrounding tissue.
  • the therapeutically effective amount is such to produce a dose of light activated drug within a range of from about 0.1 to about 20 mg/kg, preferably from about
  • the w/v concentration of the light activated drug in the composition ranges from about 0.1 to about 8.0-10.0 g/L. Most preferably, the concentration is about 2.0 to 2.5 g L.
  • the hydration step should take place at a temperature that does not exceed about
  • the glass transition temperature of the light activated drag-lipid complex can be measure by using a differential scanning microcalorimeter.
  • the lipid membrane tends to undergo phase transition from a "solid" gel state to a pre- transition state and, finally, to a more "fluid” liquid crystal state.
  • these higher temperatures however, not only does fluidity increase, but the degree of phase separation and the proportion of membrane defects also increases.
  • the usual thickness of a lipid bilayer in the "solid" gel state decreases in the transition to the "liquid" crystalline state to about 37 A, thus shrinking the entrapped volume available for the light activated drags to occupy.
  • the smaller "room” is not capable of containing as great a volume of light activated drag, which can then be squeezed out of the saturated lipid bilayer interstices. Any two or more liposomes exuding light activated drug may aggregate together, introducing further difficulties with respect to particle size reduction and ease of sterile filtration.
  • the use of higher hydration temperatures such as, for example, about 35 ° to 45 ° C, can also result in losses of light activated drag potency as the light activated drag either precipitates or aggregates during aseptic filtration.
  • the particle sizes of the coarse liposomes first formed in hydration are then homogenized to a more uniform size, reduced to a smaller size range, or both, to about 150 to 300 nm, preferably also at a temperature that does not exceed about 30° C, preferably below the glass transition temperature of the light activated drag-phospholipid complex formed in the hydration step, and even more preferably below room temperature of about 25 ° C.
  • Various high-speed agitation devices may be used during the homogemzation step, such as a MicrofluidizerTM model 110F; a sonicator; a high-shear mixer; a homogenizer; or a standard laboratory shaker.
  • the homogenization temperature should be at room temperature or lower, e.g., 15°-20° C.
  • the relative filterability of the liposome composition may improve initially due to increased fluidity as expected, but then, unexpectedly, tends to decrease with continuing agitation due to increasing particle size.
  • a high pressure device such as MicrofluidizerTM is used for agitation.
  • a high pressure device such as MicrofluidizerTM is used for agitation.
  • a great amount of heat is generated during the short-period of time during which the fluid passes through a high pressure interaction chamber.
  • two streams of fluid at a high speed collide with each other at a 90° angle.
  • the fluidity of the membrane increases, which initially makes particle size reduction easier, as expected.
  • filterability can increase by as much as four times with the initial few passes through a MicrofluidizerTM device.
  • the increase in the fluidity of the bilayer membrane promotes particle size reduction, which makes filtration of the final composition easier. In the initial several passes, this increased fluidity mechanism advantageously dominates the process.
  • the homogenization temperature is cooled down to and maintained at a temperature no greater than room temperature after the composition passes through the zone of maximum agitation, e.g., the interaction chamber of a MicrofluidizerTM device.
  • An appropriate cooling system can easily be provided for any standard agitation device in which homogenization is to take place, e.g., a MicrofluidizerTM, such as by circulating cold water into an appropriate cooling jacket around the mixing chamber or other zone of maximum turbulence. While the pressure used in such high pressure devices is not critical, pressures from about 10,000 to about 16,000 psi are not uncommon.
  • the compositions are preferably aseptically filtered through a filter having an extremely small pore size, i.e., 0.22 ⁇ m.
  • Filter pressures used during sterile filtration can vary widely, depending on the volume of the composition, the density, the temperature, the type of filter, the filter pore size, and the particle size of the liposomes.
  • a typical set of filtration conditions would be as follows: filfration pressure of 15-25 psi; filtration load of 0.8 to 1.5 ml/cm 2 ; and filfration temperature of about 25 ° c.
  • the liposome composition may be freeze-dried for long-term storage if desired.
  • BPD-MA a preferred light activated drag
  • the composition may be packed in vials for subsequent reconstitution with a suitable aqueous solution, such as sterile water or sterile water containing a saccharide and/or other suitable excipients, prior to admimstration, for example, by injection.
  • liposomes that are to be freeze-dried are formed upon the addition of an aqueous vehicle contain a disaccharide or polysaccharide during hydration.
  • the composition is then collected, placed into vials, freeze-dried, and stored, ideally under refrigeration.
  • the freeze-dried composition can then be reconstituted by simply adding water for injection just prior to administration.
  • the liposomal composition provides liposomes of a sufficiently small and narrow particle size that the aseptic filtration of the composition through a 0.22 ⁇ m hydrophilic filter can be accomplished efficiently and with large volumes of 500 ml to a liter or more without significant clogging of the filter.
  • a particularly preferred particle size range is below about 300 nm, more preferably below from about 250 nm. Most preferably, the particle size is below about 220 nm.
  • the concentration of the light activated drags in the liposome depends upon the nature of the light activated drag used. When BPD-MA is used for example, the light activated drag is generally incorporated in the liposomes at a concentration of about 0.10% up to 0.5% w/v. If freeze-dried and reconstituted, this would typically yield a reconstituted solution of up to about 5.0 mg/ml light activated drug.
  • the light activated drags incorporated into liposomes may be used along with, or may be labeled with, a radioisotope or other detecting means. If this is the case, the detection means depends on the nature of the label. Scintigraphic labels such as technetium or indium can be detected using ex vivo scanners. Specific fluorescent labels can also be used but, like detection based on fluorescence of the light activated drugs themselves, these labels can require prior irradiation. The methods of preparing various light activated drugs, light activated drug conjugates, emulsions and microbubbles are described in greater detail in the examples below.
  • Examples 5 and 6 describes a synthesis of an emulsion including a light activated drag.
  • Example 7 describes preparation of microbubbles which include a light activated drag.
  • Lutetium(HI) acetate hydrate can be purchased from Strem Chemicals, Inc. (Newburyport, Mass.), gadolinium(IH) acetate tetrahydrate can be purchased from Aesar/Johnson Matthey (Ward Hill, Mass.) and LZY-54 zeolite can be purchased from UOP (Des Plaines, III.).
  • Acetone, glacial acetic acid, methanol, ethanol, isopropyl alcohol, and n-heptanes can be purchased from J. T. Baker (Phillipsburg, N.J.).
  • Triethylamine and Amberlite 904 anion exchange resin can be purchased from Aldrich (Milwaukee, Wise). All chemicals should be ACS grade and used without further purification.
  • Figure 22 illusfrates the synthesis of the gadolinium (El) complex of 4,5-diethyl- 10,23-dimethyl-9,24-bis(3-hydroxypropyl)-16,17-bis[2-[2-(2- methoxyethoxy) ethoxy]ethoxy]-pentaazapentacyclo [20.2. l.l 3,6 .1 8 ' " .0 14 ' 19 ]heptacosa- 1,3,5,7,9,1 l(27),12,14,16,18,20,22(25),23-tridecaene which is illusfrated as Formula I of Figure 22.
  • the critical intermediate l,2-bis[2-[2-(2-methoxyethoxy)ethoxy)ethoxy]-4,5- dinifrobenzene can be prepared according to a three-step synthetic process outlined in Figure 22.
  • Formula E In an oven dried 1 L roundbottom flask Formula D (104 g, 0.26 mol) and glacial acetic acid (120 mL) are combined and cooled to 5° C. To this well stirred solution, concentrated nitric acid (80 mL) is added dropwise over 15-20 min. The temperature of the mixture is held below 40° C. by cooling and proper regulation of the rate of addition of the acid. After addition, the reaction is allowed to stir for an additional
  • the aqueous layer is extracted with CH2C12 (2x150 mL) and the combined organic extracts washed with 10% NaOH (2X250 mL) and brine (250 mL), dried (MgS0 4 ), and concentrated under reduced pressure.
  • the resulting orange oil is dissolved in acetone (100 mL), and the solution layered with n-hexanes (500 mL), and stored in the freezer.
  • Formula I 1 ,3,5,7,9, 11 (27), 12, 14, 16, 18,20,22(25),23-tridecaene, Formula I.
  • Formula I is prepared according to the process outlined in Figure 22.
  • Formula H 33.0 g, 0.036 mol
  • gadolinium(II) acetate tetrahydrate (15.4 g, 0.038 mol) are combined in methanol (825 mL).
  • methanol 825 mL
  • gadolinium(i ⁇ ) acetate tetrahydrate (15.4 g, 0.038 mol) and triethylamine (50 mL) are added and the reaction is heated to reflux.
  • the crade complex (35 g) is dissolved in MeOH (600 mL), stirred vigorously for 15 min, filtered through Celite, and transferred to a 2 L Erlenmeyer flask. An additional 300 mL of MeOH and 90 mL water are added to the flask, along with acetic acid washed LZY-54 zeolite (150 g). The suspension is agitated with an overhead mechanical stirrer for approximately 3-4 h. The zeolite extraction is deemed complete with the absence of free
  • the zeolite is removed through a Whatman #3 filter paper and the collected solids rinsed with MeOH (200 mL).
  • the dark green filtrate is loaded onto a column of Amberlite IRA-904 anion exchange resin (30 cm length x 2.5 cm diameter) and eluted through the resin (ca. 10 mL/min flow rate) into a 2 L round bottom flask with 300 mL 1-butanol.
  • the resin is rinsed with an additional 100 mL of MeOH and the combined eluent evaporated to dryness under reduced pressure.
  • the macrocyclic ligand Formula H is oxidatively metalated using lutetium(III) acetate hydrate (9.75 g, 0.0230 mol) and triethylamine (22 mL) in air-saturated methanol (1500 mL) at reflux. After completion of the reaction (as judged by the optical spectrum of the reaction mixture), the deep green solution is cooled to room temperature, filtered through a pad of celite, and the solvent removed under reduced pressure.
  • the dark green solid is suspended in acetone (600 mL, stirred for 30 min at room temperature, and then filtered to wash away the red/brown impurities (incomplete oxidation products and excess triethylamine).
  • the crade complex is dissolved into MeOH (300 mL, stirred for -30 min, and then filtered through celite into a 1 L Erlenmeyer flask. An additional 50 mL of MeOH and 50 mL of water are added to the flask along with acetic acid washed LZY-54 zeolite (40 g). The resulting mixture is agitated or shaken for 3 h, then filtered to remove the zeolite.
  • the zeolite cake is rinsed with MeOH (100 mL and the rinse solution added to the filtrate.
  • the filtrate is first concentrated to 150 mL and then loaded onto a column (30 cm length x 2.5 cm diameter) of pretreated Amberlite IRA-904 anion exchange resin (resin in the acetate form).
  • the eluent containing the bis-acetate lutetium(III) texaphyrin complex is collected, concentrated to dryness under reduced pressure, and recrystallized from anhydrous methanol/t-butylmethyl ether to afford 11.7 g (63%) of a shiny green solid.
  • FIG. 23 illustrates the synthesis of a light activated drag conjugate.
  • the light activated drag is a texaphyrin coupled with an oligonucleotide which is complementary to a DNA site.
  • the light activated drug conjugate can bind the complementary DNA site and will cleave the site upon activation by ultrasound.
  • TLC RfH3.69, 20% methanol/CHCL 3 (streaks, turns green on plate with I 2 ).
  • the compounds are dissolved in dimethylformamide (anhydrous, 500 ⁇ L) and dicyclohexylcarbodiimide (10 mg, 48 ⁇ mol) is added.
  • the resulting solution is stirred under argon with protection from Ught for 8 h, whereupon a 110 ⁇ L aliquot is added to a solution of oligodeoxynucleotide (Formula G) (87 ⁇ mol) in a volume of 350 ⁇ L of 0.4M sodium bicarbonate buffer in a 1.6 mL Eppendorf tube. After vortexing briefly, the solution is allowed to stand for 23 h with light protection.
  • the suspension is filtered through 0.45 ⁇ m nylon microfilterfuge tubes, and the Eppendorf tube is washed with 250 ⁇ L sterile water.
  • the combined filtrates are divided into two Eppendorf tubes, and glycogen (20 mg/mL, 2 ⁇ L) and sodium acetate (3M, pH 5.4 30 ⁇ L) are added to each tube.
  • glycogen (20 mg/mL, 2 ⁇ L) and sodium acetate (3M, pH 5.4 30 ⁇ L) are added to each tube.
  • ethanol absolute, 1 mL
  • Ethanol is decanted following centrifugation, and the DNA is washed with an additional 1 mL aliquot of ethanol and allowed to air dry.
  • the pellet is dissolved in 50% formamide gel loading buffer (20 ⁇ L), denatured at 90°C. for ca.
  • the suspension is filtered through nylon filters (0.45 ⁇ m) and desalted using a Sep-pakTM reverse phase cartridge.
  • the conjugate is eluted from the cartridge using 40% acetonitrile, lyophilized overnight, and dissolved in 1 mM HEPES buffer, pH 7.0 (500 ⁇ L). The solution concentration is determined using UV/vis spectroscopy.
  • EXAMPLE 3 Synthesis of texaphyrin metal complexes with amine-, thiol- or hydroxy-linked oligonucleotides
  • Amides, ethers, and thioethers are representative of linkages which may be used for coupling site-directing molecules such as oligonucleotides to light activated drugs such as texaphyrin metal complexes as illusfrated in Figure 24.
  • OUgonucleotides or other site-directing molecules functionalized with amines at the 5'-end, the 3'-end, or internally at sugar or base residues are modified post-synthetically with an activated carboxylic ester derivative of the texaphyrin complex.
  • a bromide derivatized texaphyrin (for example, Formula C of Figure 24) will react with an hydroxyl group of an oligonucleotide to form and ether linkage between the texaphyrin linker and the oligonucleotide.
  • oligonucleotide analogues containing one or more thiophosphate or thiol groups are selectively alkylated at the sulfur atom(s) with an alkyl halide derivative of the texaphyrin complex.
  • Oligodeoxynucleotide-complex conjugates are designed so as to provide optimal catalytic interaction between the targeted DNA phosphodiester backbone and the texaphyrino.
  • Oligonucleotides are used to bind selectively compounds which include the complementary nucleotide or oligo- or polynucleotides containing substantially complementary sequences.
  • a substantially complementary sequence is one in which the nucleotides generally base pair with the complementary nucleotide and in which there are very few base pair mismatches.
  • the oligonucleotide may be large enough to bind probably at least 9 nucleotides of complementary nucleic acid.
  • oligomers up to ca. 100 residues in length are prepared on a commercial synthesizer, eg., Applied Biosystems Inc. (ABI) model 392, that uses phosphoramidite chemistry. DNA is synthesized from the 3' to the 5' direction through the sequential addition of highly reactive phosphorous(IH) reagents called phosphoramidites. The initial 3' residue is covalently attached to a controlled porosity silica solid support, which greatly facilitates manipulation of the polymer.
  • ABS Applied Biosystems Inc.
  • the phosphoras(III) is oxidized to the more stable phosphoras(V) state by a short treatment with iodine solution. Unreacted residues are capped with acetic anhydride, the 5'-protective group is removed with weak acid, and the cycle may be repeated to add a further residue until the desired DNA polymer is synthesized. The full length polymer is released from the solid support, with concomitant removal of remaining protective groups, by exposure to base.
  • a common protocol uses saturated ethanolic ammonia.
  • the phosphonate based synthesis is conducted by the reaction of a suitably protected nucleotide containing a phosphonate moiety at a position to be coupled with a solid phase-derivatized nucleotide chain having a free hydroxyl phosphonate ester linkage, which is stable to acid.
  • the oxidation to the phosphate or thiophosphate can be conducted at any point during synthesis of the oligonucleotide or after synthesis of the oligonucleotide is complete.
  • the phosphonates can also be converted to phosphoramidate derivatives by reaction with a primary or secondary amine in the presence of carbon tetrachloride.
  • a protected phosphodiester nucleotide is condensed with the free hydroxyl of a growing nucleotide chain derivatized to a solid support in the presence of coupling agent.
  • the reaction yields a protected phosphate linkage which may be treated with an oximate solution to form unprotected oligonucleotide.
  • oligonucleotides may also be synthesized using solution phase methods such as diester synthesis. The methods are workable, but in general, less efficient for oligonucleotides of any substantial length.
  • Preferred oligonucleotides resistant to in vivo hydrolysis may contain a phosphorothioate substitution at each base (J. Org. Chem. 55:4693-469, (1990) and Agrawal, (1990)). Oligodeoxynucleotides or their phosphorothioate analogues may be synthesized using an Applied Biosystem 380B DNA synthesizer (Applied Biosystems, Inc., Foster City, Calif.).
  • EXAMPLE 4 Synthesis of Diformyl Monoacid Tripyrrane ( Figure 25, Formula H) and Oligonucleotide Conjugate ( Figure 25, Formula J)
  • the present example provides for the synthesis of a light activated drag conjugate.
  • the light activated drag conjugate includes a oligonucleotide acting as a site directing molecule coupled with the tripyrrane portion of a texaphyrin as illusfrated in Figure 25.
  • a three-neck 2000 mL round-bottom flask set with a magnetic stirring bar, a hydrogen line, and a vacuum line is charged with dimethylester dibenzylester dipyrromethane (Formula B) (33.07 g, 53.80 mmol), anhydrous tetrahydrofuran (1500 mL), and 10% palladium on charcoal (3.15 g.)
  • the flask is filled with dry hydrogen gas after each of several purges of the flask atmosphere prior to stirring the reaction suspension under a hydrogen atmosphere for 24 hours.
  • the dry solids are suspended in a mixture of saturated aqueous sodium bicarbonate (1500 mL) and ethyl alcohol (200 mL), and stirred at its boiling point for five minutes.
  • the hot suspension is filtered over celite.
  • the filtrate is cooled down to room temperature and acidified to pH 6 with 12N aqueous hydrochloric acid.
  • the resulting mixture is filtered over medium fritted glass.
  • the cold mixture is filtered over medium fritted glass.
  • the collected solids are washed with hexanes and dried under high vacuum overnight (13.05 g, 19.25 mmol, 39.85 yield).
  • Methylester Diacid Tripyrrane, Formula F All the glassware is oven dried. A three-neck 500 mL round-bottom flask set with a magnetic stirring bar, a hydrogen line, and a vacuum line is charged with methylester dibenzylester tripyrrane (Formula E) (12.97 g, 19.13 mmol), anhydrous tetrahydrofuran (365 mL), and 10% palladium on charcoal (1.13 g.) The flask is filled with dry hydrogen gas after each of several purges of the flask atmosphere prior to stirring the reaction suspension for 24 hours under a hydrogen atmosphere at room temperature.
  • methylester dibenzylester tripyrrane (Formula E) (12.97 g, 19.13 mmol)
  • anhydrous tetrahydrofuran 365 mL
  • 10% palladium on charcoal (1.13 g.
  • reaction suspension is filtered over celite.
  • solvent of the filtrate is removed under reduced pressure to obtain a foam which is dried under high vacuum overnight (10.94 g, 21.99 mmol, 87.0% pure.)
  • Triethylorthoformate (32.5 mL) is dripped into the flask from the addition funnel over a 20 minute period keeping the flask contents below -25 °C by means of a dry ice/ethylene glycol bath. The reaction solution is stirred for one hour at -25 °C and then a 0°C bath is set up. Deionized water (32.5 mL) is dripped into the reaction flask from the addition funnel keeping the flask contents below 10°C. The resulting two phase mixture is stirred at room temperature for 75 minutes and then added 1-butanol (200 mL.) The solvents are removed under reduced pressure.
  • the monoacid tripyrrane (Formula H) is condensed with a derivatized ortho- phenylene diamine to form a nonaromatic precursor which is then oxidized to an aromatic metal complex, for example, Formula I.
  • An oligonucleotide amine may be reacted with the carboxylic acid derivatized texaphyrin Formula I to form the conjugate Formula J having the site-directing molecule on the T (tripyrrane) portion of the molecule rather than the B (benzene) portion.
  • MCT oil medium chain length oil
  • SnEt 2 medium chain length oil
  • Certain emulsions also included additional excipients in the following concenfrations: ethanol at mg/gm oil; egg phospholipids at 75 mg/gm oil; and sodium cholate at 10 mg/gm oil. After incubating for 30 minutes at 55° C, the tubes stand overnight at room temperature (19°- 22° C).
  • the tubes are centrifuged to remove bulk precipitates, and supematants are filtered through 0.45 ⁇ m nylon membrane to remove any undissolved drag. Aliquots of filtrate are then diluted in chloroform:isopropyl alcohol (1:1) for spectrophotometric determination of drag concentration (absorbance at 662 nm). Reference standards are prepared with known concentrations of SnEt 2 in the same solvent. The concentration of SnEt 2 in each of the emulsions is illustrated in Table 2. As illusfrated, the concenfration of SnEt 2 in the emulsion can be more than ten times the concentration in MCT oil alone.
  • MCT oil, egg phospholipids, eth.anol, .and SnEt 2 are incubated with different bile salts, all at 4.6 mM 1 , under the same conditions described above.
  • sodium cholate is the most efficient solubilizer.
  • Cholic acid lacks solubilizing action in the oil.
  • Example 1 illustrates the preparation of liposomes including BPD- MA (See Figure 17) as a light activated drug.
  • a 100-ml batch of BPD-MA liposomes is prepared at room temperature (about 20° C.) using the following general procedure.
  • BPD-MA, butylated hydroxytoluene ("BHT"), ascorbyl pahnirate, and the phospholipids DMPC and EPG are dissolved in methylene chloride.
  • the molar ratio of light activated drag: EPG:DMPC is 1.0:3.7 and has the compositions illustrated in Table 4.
  • the total lipid concenfration (% w/v) is about 2.06.
  • the resulting solution is filtered through a 0.22 ⁇ m filter and then dried under vacuum using a rotary evaporator. Drying is continued until the amount of methylene chloride in the solid residue is no longer detectable by gas chromatography.
  • a 10% lactose/water-for-injection solution is then prepared and filtered through a
  • the lactose/water solution is allowed to remain at room temperature (about 25 ° C.) for addition to the flask containing the solid residue of the light activated drug/phospho lipid.
  • the solid residue is dispersed in the 10% lactose/water solution at room temperature, stirred for about one hour, and passed through a MicrofluidizerTM homogenizer three to four times with the outlet temperature controlled to about 200°-250° C.
  • the solution is then filtered through a 0.22 ⁇ m Durapore, hydrophilic filter.
  • the filterability of the composition in g/cm 2 is typically greater than about 10. Moreover, the yield is about 100% by HPLC analysis, with light activated drug potency typically being maintained even after sterile filtration. Average particle sizes vary from about 150 to about 300 nm ( ⁇ 50 nm).
  • Example 2 describes the delivery of a light activated drag to an atheroma.
  • An emulsion is prepared having about .6 g SnEt 2 /ml of emulsion and about 20 g of MCT oil based hydrophobic phase/ml of emulsion.
  • the 7C is positioned in a vessel of the cardiovascular system using over the guidewire techniques.
  • the catheter is positioned such that the media delivery port is adjacent to the atheroma using radiopaque markers on the catheter and the balloon is expanded into contact with the vessel wall.
  • the emulsion is delivered via the third utility lumen 16B of the catheter 10. After the delivery of the emulsion, the ultrasound energy is delivered at about 0.3 W/cm 2 at a frequency of approximately 1.3 MHz for about ten minutes. After the delivery of ulfrasound energy has concluded, the catheter is withdrawn from the vasculature of the tumor.
  • the following Example describes the delivery of a light activated drag to a tumor.
  • An emulsion is prepared having approximately .8 g SnEt 2 /ml of emulsion and approximately 30 g of MCT oil based hydrophobic phase/ml of emulsion.
  • the catheter 10 illustrated in Figure 3 A is positioned in the vasculature of a rumor using over the guidewire techniques.
  • the catheter is positioned such that the media delivery port is within the tumor using radiopaque markers included on the catheter.
  • the prepared emulsion is delivered into the vasculature of the tumor via the utility lumen 16 A.
  • the ulfrasound energy is delivered at about 0.3 W/cm 2 at a frequency of approximately 1.3 MHz for about fifteen minutes.
  • the catheter is withdrawn from the vascular system of the patient.
  • Example 2 describes the delivery of a light activated drug to a potential restenosis site.
  • An emulsion is prepared having approximately .6 g SnEt 2 /ml of emulsion and approximately 30 g of MCT oil based hydrophobic phase/ml of emulsion.
  • the catheter illusfrated in Figure 7C is positioned in the vasculature of a patient using over the guidewire techniques.
  • the catheter is positioned such that the media delivery port is adjacent to a portion of the vessel which was previously treated with balloon angioplasty and the balloon is expanded into contact with the vessel wall.
  • the prepared emulsion is delivered into the vasculature of the patient via the third utility lumen 16B.
  • Ultrasound energy is delivered from the ultrasound assembly to the potential restenosis site at about 0.3 W/cm 2 at a frequency of approximately 1.3 MHz for about ten minutes. After the delivery of ultrasound energy has concluded, the catheter is withdrawn from the vascular system of the patient.
  • MA:EPG:DMPC is about 1:3:7.
  • the catheter illusfrated in Figure 7C is positioned in a vessel of the cardiovascular system using over the guidewire techniques.
  • the catheter is positioned such that the media delivery port is adjacent to the atheroma using radiopaque markers included on the catheter and the balloon is expanded into contact with the vessel.
  • Ultrasound energy is delivered at about 0.3 W/cm 2 at a frequency of approximately 1.3
  • the catheter is withdrawn from the vascular system of the patient.
  • Liposomes are prepared including BPD-MA (See Figure 17) as the light activated drug and DMPC and EPG as the phospholipids.
  • the molar ratio of BPD-MA:EPG:DMPC is about 1:3:7.
  • the catheter illustrated in Figure 8 is positioned in the vasculature of a tumor using over the guidewire techniques. The catheter is positioned such that the media delivery port is within the tumor using radiopaque markers included on the catheter. Ultrasound energy is delivered at about 0.3 W/cm 2 at a frequency of approximately 1.3
  • the catheter is withdrawn from the vasculature of the tumor.
  • Liposomes are prepared including BPD-MA (See Figure 17) as the light activated drug and DMPC and EPG as the phospholipids.
  • BPD-MA See Figure 17
  • EPG the phospholipids.
  • the molar ratio of BPD-MA:EPG:DMPC is approximately 1:3:7.
  • the catheter illusfrated in Figure 7C is positioned in the vasculature of a patient using over the guidewire techniques. The catheter is positioned such that the media delivery port is adjacent to a portion of the vasculature which was previously treated with balloon angioplasty and the balloon is inflated into contact with the vessel wall.
  • Ultrasound energy is delivered at about 0.3 W/cm 2 at a frequency of approximately 1.3 MHz for about 15 minutes in order to rapture the liposomes and cause tissue death within the atheroma. After the delivery of ultrasound energy is concluded, the catheter is withdrawn from the vasculature of the patient.
  • the following Example describes the delivery of a light activated drag to an atheroma.
  • Liposomes are prepared including BPD-MA (See Figure 17) as the light activated drug and DMPC and EPG as the phospholipids.
  • the molar ratio of BPD- MA:EPG:DMPC is about 1:3:7.
  • the phospholipids are systemically delivered.
  • the catheter illustrated in Figure 7C is positioned in the vasculature of a patient using over the guidewire techniques. The catheter is positioned such that the media delivery port is adjacent to the atheroma and the balloon is inflated into contact with the vessel wall.
  • Ultrasound energy is delivered at about 0.3 W/cm 2 at a frequency of approximately 1.3 MHz for about 15 minutes. After the delivery of ultrasound energy is concluded, the catheter is withdrawn from the vasculature of the patient.
  • Example 2 describes the delivery of a light activated drag to a tumor.
  • Microbubbles are prepared including cisplatin and photofrin according to the methods disclosed in U.S. patent number 5,770,222.
  • the microbubbles are systemically administered.
  • the catheter illustrated in Figure 1 A is positioned within the vasculature of a tumor. Ulfrasound energy is delivered at about 0.3 W/cm 2 at a frequency of approximately 1.3 MHz for about 15 minutes. After the delivery of ultrasound energy is concluded, the catheter is withdrawn from the vasculature of the patient.
  • Example 2 describes the delivery of a light activated drag to a tumor.
  • Microbubbles .are prepared including cisplatin and photofrin according to the methods disclosed in U.S. patent number 5,770,222.
  • the catheter illustrated in Figure 3A is positioned within the vasculature of a tumor.
  • the microbubbles .are delivered to the tumor via the second utility lumen 16A of the catheter.
  • Ulfrasound energy is delivered at about 0.3 W/cm 2 at a frequency of approximately 1.3 MHz for about 15 minutes. After the delivery of ulfrasound energy is concluded, the catheter is withdrawn from the vasculature of the patient.
  • Example 2 describes the delivery of a light activated drag to a thrombosis.
  • Microbubbles are prepared including heparin, photofrin and an albumin subsfrate.
  • the microbubbles are systemically administered.
  • the catheter illustrated in Figure 1 A is positioned adjacent to the thrombosis. Ulfrasound energy is delivered at about 0.2 W/cm 2 at a frequency of approximately 1.3 MHz for about 20 minutes. After the delivery of ultrasound energy is concluded, the catheter is withdrawn from the vasculature of the patient.

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Abstract

On décrit un kit et un procédé utilisé pour provoquer la mort des tissus à l'intérieur d'un site tissulaire. Le kit comprend un support comportant un médicament activé par la lumière qui lors de l'exposition à un niveau spécifique d'énergie ultrasonore peut être activé. Le kit comprend également un cathéter dont une lumière est reliée à un orifice d'apport du support à travers laquelle le médicament activé par la lumière peut être apporté localement au site tissulaire. Le transducteur ultrasonore est configuré pour envoyer le niveau d'énergie ultrasonore qui active le médicament activé par la lumière avec une puissance suffisante pour que l'énergie ultrasonore puisse pénétrer le site tissulaire.
PCT/US1998/019797 1996-03-05 1998-09-21 Ensemble a ultrasons destine a etre utilise avec des medicaments actives par la lumiere WO1999013943A1 (fr)

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AU95005/98A AU9500598A (en) 1997-09-19 1998-09-21 Ultrasound assembly for use with light activated drugs

Applications Claiming Priority (7)

Application Number Priority Date Filing Date Title
US61110596A 1996-03-05 1996-03-05
JP25581497A JP4791616B2 (ja) 1997-09-19 1997-09-19 薬物坦持体及びその使用方法
JP9/255814 1997-09-19
US97284697A 1997-11-18 1997-11-18
US08/972,846 1997-11-18
US09/129,980 1998-08-05
US09/129,980 US6210356B1 (en) 1998-08-05 1998-08-05 Ultrasound assembly for use with a catheter

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1180380A2 (fr) * 2000-08-17 2002-02-20 William N. Borkan Cathéter pour traitement médical
WO2001085213A3 (fr) * 2000-05-08 2002-08-01 Univ British Columbia Supports pour formulations photosensibilisantes
US7131963B1 (en) 2002-06-27 2006-11-07 Advanced Cardiovascular Systems, Inc. Catheters and methods of using catheters
US7267659B2 (en) 2002-05-24 2007-09-11 Dornier Medtech Systems Gmbh Method and apparatus for transferring medically effective substances into cells
WO2012143739A1 (fr) * 2011-04-21 2012-10-26 University Of Ulster Thérapie sonodynamique
US9060915B2 (en) 2004-12-15 2015-06-23 Dornier MedTech Systems, GmbH Methods for improving cell therapy and tissue regeneration in patients with cardiovascular diseases by means of shockwaves
EP3117784A1 (fr) * 2009-07-08 2017-01-18 Sanuwave, Inc. Utilisation d'ondes de choc de pression intracorporelle en médecine

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WO1991009629A1 (fr) * 1989-12-22 1991-07-11 Unger Evan C Liposomes utilises comme agents de contraste pour imagerie ultrasonique
WO1994005361A1 (fr) * 1992-08-28 1994-03-17 Cortrak Medical, Inc. Appareil a matrice polymere d'administration de medicament et procede
JPH0748710A (ja) 1994-01-10 1995-02-21 Yoshiaki Kakine 天然草帽子
WO1996027341A1 (fr) * 1995-03-08 1996-09-12 Ekos, Llc Appareil a ultrasons a usage therapeutique
WO1996035469A1 (fr) * 1995-05-10 1996-11-14 Cardiogenesis Corporation Systeme de traitement ou de diagnostic pour le tissu cardiaque
WO1996036286A1 (fr) * 1995-05-15 1996-11-21 Coraje, Inc. Thrombolyse a ultrason amelioree
US5578291A (en) 1993-05-14 1996-11-26 The Board Of Regents Of The University Of Nebraska Method and composition for optimizing left ventricular videointensity in echocardiography
US5664382A (en) 1993-09-09 1997-09-09 Melnick; David W. Method for making block forms for receiving concrete
US5701899A (en) 1993-05-12 1997-12-30 The Board Of Regents Of The University Of Nebraska Perfluorobutane ultrasound contrast agent and methods for its manufacture and use
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Publication number Priority date Publication date Assignee Title
US4883790A (en) 1987-01-20 1989-11-28 University Of British Columbia Wavelength-specific cytotoxic agents
US4920143A (en) 1987-04-23 1990-04-24 University Of British Columbia Hydro-monobenzoporphyrin wavelength-specific cytotoxic agents
WO1991009629A1 (fr) * 1989-12-22 1991-07-11 Unger Evan C Liposomes utilises comme agents de contraste pour imagerie ultrasonique
US5770222A (en) * 1989-12-22 1998-06-23 Imarx Pharmaceutical Corp. Therapeutic drug delivery systems
WO1994005361A1 (fr) * 1992-08-28 1994-03-17 Cortrak Medical, Inc. Appareil a matrice polymere d'administration de medicament et procede
US5701899A (en) 1993-05-12 1997-12-30 The Board Of Regents Of The University Of Nebraska Perfluorobutane ultrasound contrast agent and methods for its manufacture and use
US5578291A (en) 1993-05-14 1996-11-26 The Board Of Regents Of The University Of Nebraska Method and composition for optimizing left ventricular videointensity in echocardiography
US5664382A (en) 1993-09-09 1997-09-09 Melnick; David W. Method for making block forms for receiving concrete
JPH0748710A (ja) 1994-01-10 1995-02-21 Yoshiaki Kakine 天然草帽子
WO1996027341A1 (fr) * 1995-03-08 1996-09-12 Ekos, Llc Appareil a ultrasons a usage therapeutique
WO1996035469A1 (fr) * 1995-05-10 1996-11-14 Cardiogenesis Corporation Systeme de traitement ou de diagnostic pour le tissu cardiaque
WO1996036286A1 (fr) * 1995-05-15 1996-11-21 Coraje, Inc. Thrombolyse a ultrason amelioree

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001085213A3 (fr) * 2000-05-08 2002-08-01 Univ British Columbia Supports pour formulations photosensibilisantes
EP1180380A3 (fr) * 2000-08-17 2003-01-22 William N. Borkan Cathéter pour traitement médical
EP1180380A2 (fr) * 2000-08-17 2002-02-20 William N. Borkan Cathéter pour traitement médical
US7267659B2 (en) 2002-05-24 2007-09-11 Dornier Medtech Systems Gmbh Method and apparatus for transferring medically effective substances into cells
US7131963B1 (en) 2002-06-27 2006-11-07 Advanced Cardiovascular Systems, Inc. Catheters and methods of using catheters
US9060915B2 (en) 2004-12-15 2015-06-23 Dornier MedTech Systems, GmbH Methods for improving cell therapy and tissue regeneration in patients with cardiovascular diseases by means of shockwaves
US10058340B2 (en) 2009-07-08 2018-08-28 Sanuwave, Inc. Extracorporeal pressure shock wave devices with multiple reflectors and methods for using these devices
EP3117784A1 (fr) * 2009-07-08 2017-01-18 Sanuwave, Inc. Utilisation d'ondes de choc de pression intracorporelle en médecine
US10238405B2 (en) 2009-07-08 2019-03-26 Sanuwave, Inc. Blood vessel treatment with intracorporeal pressure shock waves
US10639051B2 (en) 2009-07-08 2020-05-05 Sanuwave, Inc. Occlusion and clot treatment with intracorporeal pressure shock waves
US11666348B2 (en) 2009-07-08 2023-06-06 Sanuwave, Inc. Intracorporeal expandable shock wave reflector
US11925366B2 (en) 2009-07-08 2024-03-12 Sanuwave, Inc. Catheter with multiple shock wave generators
US12004760B2 (en) 2009-07-08 2024-06-11 Sanuwave, Inc. Catheter with shock wave electrodes aligned on longitudinal axis
US12004759B2 (en) 2009-07-08 2024-06-11 Sanuwave, Inc. Catheter with shock wave electrodes aligned on longitudinal axis
US12239332B2 (en) 2009-07-08 2025-03-04 Sanuwave, Inc. Catheter with multiple shock wave generators
WO2012143739A1 (fr) * 2011-04-21 2012-10-26 University Of Ulster Thérapie sonodynamique

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