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WO1988001509A1 - Compositions pharmacologiquement actives de butanes catecholiqueset de zinc - Google Patents

Compositions pharmacologiquement actives de butanes catecholiqueset de zinc Download PDF

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Publication number
WO1988001509A1
WO1988001509A1 PCT/US1986/001740 US8601740W WO8801509A1 WO 1988001509 A1 WO1988001509 A1 WO 1988001509A1 US 8601740 W US8601740 W US 8601740W WO 8801509 A1 WO8801509 A1 WO 8801509A1
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WO
WIPO (PCT)
Prior art keywords
composition
zinc
bis
catecholic butane
dimethylbutane
Prior art date
Application number
PCT/US1986/001740
Other languages
English (en)
Inventor
Russell T. Jordan
Larry M. Allen
Original Assignee
Chemex Pharmaceuticals, Inc.
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
Application filed by Chemex Pharmaceuticals, Inc. filed Critical Chemex Pharmaceuticals, Inc.
Priority to PCT/US1986/001740 priority Critical patent/WO1988001509A1/fr
Publication of WO1988001509A1 publication Critical patent/WO1988001509A1/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/28—Compounds containing heavy metals
    • A61K31/315—Zinc compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/24—Heavy metals; Compounds thereof
    • A61K33/30—Zinc; Compounds thereof

Definitions

  • This invention relates to pharmacologically active compositions of catecholic butanes and ionic zinc and to the use thereof in the treatment of diseases and disorders of the skin, and to their use as antibacterial and antifungal agents. They are also useful in the treatment of benign, premalignant and malignant solid tumors, especially those of the skin.
  • Mammals can be affected with a wide variety of skin disorders including bacterial and fungal infections and benign, premalignant and malignant growths.
  • the systemic application of antibiotics has been commonly used as a treatment for bacterial infections in conditions such as acne and osteomyelitis. More recently, the topical use of certain antibiotics has been reported. For example, erythromycin in combination with zinc acetate has been reported as being useful in the topical treatment of acne. Many other chemicals have been reported as having antimicrobial activity. Some of these, such as the meso form of nordihydroguaiaretic acid [meso-1,4-bis (3 ,4-dihydroxyphenyl)-2,3-dimethylbutane] which occurs naturally, have been used as folk remedies.
  • NDGA is used herein to refer to the meso form of nordihydroguaiaretic acid.
  • NDGA is a principal component in the creosote bush which has been used to make a tea used as a folk remedy for colds, rheumatism and other ailments for centuries.
  • most of these chemicals including NDGA have not proven to be successful in the treatment of conditions such as acne and osteomyelitis.
  • the general structure for nordihydroguaiaretic acid is criven in Formula (I).
  • nordihydroguaiaretic acid in a pharmaceutical composition containing ionic zinc is effective in treating disorders of the skin including bacterial infections which occur in acne and in osteomyelitis when applied to the situs of the disorder.
  • Such compositions are also effective in treating benign, premalignant and malignant growths of the skin without the detrimental side effects associated with chemosurgical techniques, when applied topically to or injected into the growth.
  • compositions comprising a catecholic butane of formula
  • R 1 and R 2 are independently H, C 1 -C 6 alkyl, or a
  • R 3 and R 4 are independently H, CH 3 , or C 2 H 5 ;
  • R 5 and R 6 are independently H or OH
  • R 7 , R 8 and R 9 may be attached at any separate position
  • C 2 -C 6 of the benzene ring and are independently H, OH, or OR 1 (wherein R 1 is as above); and a pharmaceutically acceptable source of ionic zinc.
  • this invention relates to pharmaceutical compositions adapted for topical administration comprising, in admixture with a pharmaceutically acceptable carrier, a mixture of (i) a catecholic butane of Formula (II) and (ii) a pharmaceutically acceptable source of ionic zinc.
  • this invention relates to such compositions, adapted for topical application to a situs or injection into the interior of the situs, comprising nordihydroguaiaretic acid and a source of ionic zinc, and to such compositions in combination with a pharmaceutically acceptable carrier.
  • this invention relates to a method for inhibiting the proliferation of abnormal cells in a mammal which comprises applying directly to the situs of the abnormal cells an amount of a composition of this invention effective to inhibit said proliferation.
  • this invention relates to a method of promoting the healing of a lesion in the tissue of a mammal which comprises applying thereto an amount of a composition according to this invention effective to promote the healing thereof.
  • this invention relates to a method of increasing the oxidative stability of a catecholic butane of Formula (II) which comprises mixing with said catecholic butane an oxidation inhibiting amount of ionic zinc .
  • this invention relates to a method of enhancing the retention time of a catecholic butane at the situs of an affliction to which said catecholic butane is applied, which comprises applying said catecholic butane as a composition according to this invention containing an amount of said source of ionic zinc effective to enhance said retention time.
  • source of ionic zinc means a compound comprising ionic zinc in salt or chelated form, as opposed to metallic zinc. That source can be or can include the catecholic butane itself.
  • zinc means zinc in its ionic or divalent state and not metallic zinc.
  • afflicted situs or “situs” as used herein refer to a localized area of pathology, infection, lesion or wound, or abnormal cells including solid tumors, and the immediately surrounding area.
  • applying as used herein embraces both topical applications to a surface of the situs and injection into the interior of the situs.
  • mammal as used herein includes feline, canine, equine, bovine, rodent and primate species, including cats, dogs, horses, rats, mice, monkeys and humans. Other animals, e.g., birds, can also be successfully treated with the compositions of this invention.
  • abnormal cells embraces both benign, premalignant and malignant cells. Examples of the former include the cells associated with adenomas, papillomas, etc. Examples of premalignant cells include actinic keratosis.
  • proliferation refers to the reproduction or multiplication and growth of cells.
  • escharotic means a corrosive or caustic agent which is capable of killing healthy, living cells .
  • concentration means a concentration of the source of ionic zinc which does not kill living cells upon contact, e.g., as does zinc chloride when employed as an escharotic agent, e.g., at a concentration of about 40 weight percent or higher, depending on the delivery vehicle.
  • compositions comprising a catecholic butane and zinc are particularly effective for the treatment of a variety of skin disorders and solid tumors. Improved results are obtained when the affected area is directly contacted with the instant compositions. With such compositions it has also been found that, surprisingly, the catecholic butane is retained by the tissue at the treatment site for a significant period of time before being distributed throughout the organism. This unexpected property of the instant compositions can increase the effectiveness of the treatment and also minimize any detrimental side effects of the components. Additionally, the combination of a catecholic butane and zinc allows a reduction in the concentration of each individual component while maintaining the efficacy of the composition. This reduction in the dosage level of the individual components obtained by combining the two active ingredients increases the safety of the composition.
  • compositions have been found to unexpectedly provide improved restoration of integrity to injured tissue.
  • the presence of zinc has also been found to substantially increase the stability of the catecholic butane to oxidative reactions.
  • the catecholic butane and zinc also unpredictably show no evidence of pharmacological antagonism.
  • novel compositions of this invention are useful as antimicrobial, antifungal, antiviral and antitumor agents, as lesion healing promoting agents, e.g. for skin ulcers such as decubitus ulcers and lesions associated with osteomyelitis. They are useful in the treatment of keratoses, especially actinic keratosis including senile keratotic lesions. They are useful in treating a wide variety of premalignant and malignant skin tumors, basal cell carcinoma, squamous cell carcinoma and a diversified variety of melanotic lesions which are premalignant or malignant as well as certain cutaneous tumor manifestations of otherwise systemic diseases.
  • compositions have been found to be effective against solid tumors arising from all three embryonic tissue types, namely squamous cell carcinoma, e.g. lung carcinoma, arising from the ectodermal layer; adenocarcinomas, e.g. breast, renal and colon cancer, arising from the endodermal layer; and melanoma and brain cancers, arising from the mesodermal layer.
  • the catecholic butanes useful in the compositions of the instant invention are of the Formula (II)
  • R 1 and R 2 are independently H, C 1 -C 6 alkyl, or C 7 or lower acyl;
  • R 3 and R 4 are independently H, CH 3 , or C 2 H 5 ;
  • R 5 and R 6 are independently H or OH
  • R 7 R 8 and R 9 may be attached at any separate position
  • C 2 _ C 6 of the benzene ring and are independently H, OH, or
  • Illustrative classes of compounds within the scope of Formula (II) are those wherein: a) one or more of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 and R 9 are H, e.g., those wherein R 5 is H, R 5 and R 6 are
  • R 5 , R 6 and R 7 are H and R 8 and R 9 are OH or OR 1 ;
  • R 3 and R 4 each are CH 3 or C 2 H 5 including those of a), especially those wherein R 5 , R 6 and R 7 are H and/or R 8 and R 9 are OH or OR 1 ;
  • R 1 and R 2 are acyl, e.g.. hydrocarbonacyl, preferably, alkanoyl, e.
  • R 1 and R 2 are alike and R 8 and R 9 are OR 1 , including those of a), b) and c); and e)
  • the compound is in the form of a single optical isomer, a mixture of such isomers, e.g., a racemic mixture, or a mixture of diastereoisomers, including each of a), b), c) and d).
  • alkyl represents, inter alia, methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, tert-butyl, n-pentyl, isopentyl, n-hexyl, etc.
  • Acyl represents groups having the general formula RCO-, e.g., acetyl (CH 3 CO-), propionyl (CH 3 CH 2 CO-), butyryl (CH 3 CH 2 CH 2 CO-), 3,3-dimethylbutyryl [(CH 3 )3CCH 2 CO-], etc.
  • the catecholic butane compound is named as a substituted phenyl, the corresponding groups are acetoxy (CH 3 CO 2 -), propionyloxy (CH 3 CH 2 CO 2 -), and butyryloxy (CE 3 ,C H2 CH 2 CO 2 -), 3,3-dimethylbutyryloxy [(CH 3 ) 3 CCH 2 CO 2 -], etc.
  • catecholic butanes include the d-, 1-, racemic mixture of d- and 1-, and meso-isomers of 1,4- bis(3,4-dihydroxyphenyl)-2,3-dimethylbutane; 1,4-bis(3,4- dihydroxyphenyl)butane; 1,4-bis(3,4-dimethoxyphenyl)-2,3- dimethylbutane; 1,4-bis(3,4-diethoxyphenyl)-2,3-dimethyl- butane; 1,4-bis(3,4-di ⁇ ropoxyphenyl)-2,3-dimethylbutane; 1-(3,4-dihydroxyphenyl)-4-(3',4',5'-trihydroxyphenyl) butane; 1,4-bis(3,4-diacetoxyphenyl)-2,3-dimethylbutane; 1,4-bis(3,4-dipropionyloxyphenyl)-2,3-dimethylbutane; 1,4-
  • the zinc is present in the instant compositions as a cation, e.g., as a salt or a chelate of the catecholic butane itself or as pharmaceutically acceptable salt of another toxicologically acceptable anion, or as a mixture thereof.
  • Pharmaceutically acceptable salts include those of inorganic acids, e.g., nitrate, sulfate, acetate, halides and phosphates, and those of organic acids, e.g., acetate, benzoate, citrate, caprylate, gluconate, etc., and mixtures thereof.
  • Zinc chloride is especially preferred.
  • the term "zinc" as used herein means ionic zinc, rather than zinc metal.
  • the molar ratio of catecholic butane to zinc in the compositions of this invention can vary over a wide range. Ordinarily the molar ratio is between 100 : 1 and 1 : 100 , more commonly between about 10:1 and 1:20 and even more commonly between about 5:1 and 1:15, and most commonly between about 3:1 and 1:10.
  • the molar ratio preferably is such that at least one of the catecholic butane and zinc is present in the composition at a concentration effective to inhibit the proliferation of abnormal cells and the other of the two is present therein at a concentration effective to enhance that proliferation inhibiting activity, when the composition is applied in effective amounts to the situs of those cells.
  • the molar ratio can be any that results in the composition exhibiting one or more of antifungal, antibacterial, antiviral and anti-tumor activity.
  • the zinc is present at a concentration which prolongs the half-life of the catecholic butane at the situs of application.
  • the zinc is present at a concentration which inhibits oxidation of the catecholic butane.
  • the catecholic butane and zinc are present in concentrations to promote healing of a wound or lesion contacted therewith. The preferred molar ratio range depends upon the particular condition being treated as well as the method of delivery of the composition to the treatment site.
  • the preferred range can be determined by normal pharmacological screening methods used in the art such as against the particular bacteria strain or strain of tumor cells. If desired, an excess of the zinc or the catecholic butane can be used as appropriate for the specific condition being treated.
  • the instant compositions can be applied topically to or injected into the treatment site, e.g., solid tumor, lesion or wound.
  • the catecholic butane and the source of ionic zinc are usually formulated with a pharmaceutically-acceptable carrier.
  • pharmaceutically-acceptable carrier refers to a material that is nontoxic, generally inert and does not adversely affect the functionality of the active ingredients.
  • Carrier materials are well known in the pharmaceutical formulation art and include those materials referred to as diluents or vehicles.
  • the carrier can be an inorganic or organic material and should have sufficient viscosity to allow spreading of the composition and provide good adherence to the membrane to which it is topically applied.
  • examples of such carriers include without limitation polyols such as glycerol, propylene glycol, polyethylene glycol, preferably of a molecular weight between about 400 and about 8000, suitable mixtures thereof, vegetable oils, etc.
  • the viscosity of the formulation can be adjusted by methods well known in the art, for example by the use of a higher molecular weight polyethylene glycol.
  • the formulation can contain pharmacologically-acceptable additives or adjuvants such as antimicrobial agents, e.g. methyl, ethyl, propyl, and butyl esters of para-hydroxybenzoic acid, as well as chlorobutanol, phenol, ascorbic acid, etc.
  • the formulation can also contain thickening or gelling agents, emulsifiers, wetting agents, coloring agents, buffers, stabilizers and preservatives including antioxidants such as butylhydroxyanisole.
  • the formulation can also contain penetration enhancers such as dimethyl sulfoxide, longchain alcohols such as nonoxynol, long-chain carboxylic acids, propylene glycol, N-(2-hydroxyethyl) pyrrolidone, 1-dodecyl-azacycloheptan-2-one, and the like.
  • penetration enhancers such as dimethyl sulfoxide, longchain alcohols such as nonoxynol, long-chain carboxylic acids, propylene glycol, N-(2-hydroxyethyl) pyrrolidone, 1-dodecyl-azacycloheptan-2-one, and the like.
  • absorption-delaying agents such as aluminum monostearate and gelatin.
  • composition of the formulation can be adjusted using components well-known in the formulation art to provide a pharmaceutical formulation which is a gel, cream, ointment, solid, liquid, semi-solid, etc.
  • the particular physical form of the formulation depends on the desired method of treatment and the patient to be treated.
  • the composition is formulated as a solution or suspension having a low enough viscosity to be injected.
  • the composition suitable for injectable use must be sterile and fluid to the extent that easy syringe injection exists. It should also be stable under conditions of manufacture and storage and be preserved against contamination by microorganisms. Additionally, the pH of the composition must be within a range which does not result in tissue damage.
  • the concentrations of the catecholic butane and the ionic zinc in a particular formulation depend on the condition being treated, the method of application, i.e. topical or injection, the rate of delivery of the active ingredient (s) to the treatment site, and the number of applications of the formulation which can be used. Additionally, certain catecholic butane compounds are more effective in treating particular conditions than are other analogs. The optimum amount of a specific catecholic butane for treating a condition cannot be predicted at this time. However, an effective range can readily be determined by procedures known to those skilled in the art and explained elsewhere herein. It has been found that ordinarily a lower concentration of catecholic butane and ionic zinc can be used when treating a microbial infection than when treating a solid tumor.
  • the concentration of ionic zinc in the formulation can likewise depend upon the condition being treated and the particular catecholic butane or combination of butanes being used. As discussed hereinabove, it may be desirable to have a substantial excess of one component, for example ionic zinc, present in the formulation in order to effectively treat the particular condition.
  • a formulation contain the lowest concentrations of catecholic butane and ionic zinc which effectively treat the condition with the desired number of applications, i.e. a lower effective dose rate can be tolerated if multiple applications are used.
  • This low concentration limit is dependent upon the delivery effectiveness of the carrier vehicle.
  • the catecholic butane and zinc together comprise between about 0.5 and about 80 weight percent of the formulation. Recognizing that it may be possible to use lower concentrations depending on the delivery of the carrier, it is expected that a formulation for treating microorganisms or fungi would ordinarily contain between about 0.001 and about 20 weight percent of catecholic butane and between about 0.001 and about 30 weight percent zinc.
  • the formulation contain between about 0.1 and about 30 weight percent catecholic butane and between about 0.05 and about 35 weight percent zinc.
  • at least one of the catecholic butane and the zinc is present in the formulation at a concentration of at least about 0.5 weight percent, more preferably at least about 1.0 weight percent.
  • the weight percent in the formulations refer to the concentrations of materials being effectively delivered to the treatment site.
  • formulations can be prepared that have significantly higher concentrations of catecholic butanes and zinc depending upon the carrier and additives being used.
  • the concentrations of these materials in the formulation can be substantially increased and in fact may have to be substantially increased in order to provide an effective treatment.
  • concentrations of active ingredients in a particular formulation required to provide a particular effective dose (ED) can be generally determined by a person skilled in the pharmaceutical formulation art based upon the properties of a carrier and the particular additives introduced into the formulation. It is also expected that a formulation which is being applied topically can contain a higher concentration of catecholic butane and zinc than a composition being injected, for example into a solid tumor.
  • a preferred embodiment of the instant invention comprises compositions containing nordihydroguaiaretic acid, i.e. 1,4-bis(3,4-dihydroxylphenyl)-2,3-dimethyl- butane, and zinc chloride. This combination has been found to be particularly effective in treating acne and
  • Propionibacterium acnes, decubitus ulcers, osteomyelitis, actinic keratosis and solid tumors since zinc chloride at high concentrations is an escharotic material, it is preferred that the concentration of zinc chloride delivered to the treatment site be maintained below a concentration which is escharotic to the healthy tissue.
  • the effective concentration of zinc chloride as well as nordihydroguaiaretic acid delivered to the treat ment site depends upon the carrier and other additives included in the formulation, ordinarily the concentration of nordihydroguaiaretic acid in the formulation will range from about 0.01 to about 40 weight percent and the concentration of zinc chloride in the formulation will range from about 0.01 to about 35 weight percent. These ranges are provided by way of description and not by way of limitation since it is recognized that the concentration can be adjusted over a wide range depending on the carrier material , number of applications used , etc. , as described hereinabove.
  • compositions have the advantage of the beneficial and unexpected interaction between the catecholic butane and ionic zinc.
  • This beneficial relationship is not understood at this time; but it allows the concentrations of the catecholic butane and zinc to be reduced to lower, more toxicologically acceptable levels while obtaining comparable or superior results to the use of higher concentrations. of individual components.
  • the concentration of zinc chloride can be reduced to below an escharotic level in the formulation.
  • the pH of the formulation can be important in assuring stability of the catecholic butane as well as assuring that the formulation is physiologically acceptable to the patient.
  • Many of the catechols, particularly nordihydroguaiaretic acid are susceptible to oxidation, for example by air. Such oxidation can result in discoloration of the formulation rendering it unacceptable for pharmaceutical use.
  • These catechols are more stable against oxidation at lower pH levels. Therefore, it is preferred that if the formulation is to be exposed to oxidizing conditions the pH be maintained below about 7 and preferably below about 6 in order to provide maximum stability for the catechol against oxidation. However, if oxidizing conditions can be avoided, for example by storage of the formulation under an inert atmosphere such as nitrogen, a higher pH can be used.
  • the pH of the formulation can be maintained through the use of toxicologically-acceptable buffers.
  • buffers are well known in the pharmaceutically formulation art. It has been found that the presence of ionic zinc in a catecholic butane formulation can substantially retard the rate of oxidation of the catechol, i.e. increase the stability of the catecholic butane to oxidation. This has significant advantages in that the introduction of unknown oxidation products of the catecholic butanes is minimized and the shelf-life of the catecholic butane compositions is increased.
  • ionic zinc in a molar ratio zinc to catecholic butane of about 1:50 can increase the stability of the catechol; however, it is preferred that the molar ratio zinc to catechol be at least about 1:5, and most preferably at least about 1:2 with an excess of zinc contemplated as being most beneficial.
  • compositions of the instant invention have also been found to be useful in the treatment of lesions, draining lesions, and draining wounds which show impaired healing.
  • lesion refers to any pathological or traumatic discontinuity of tissue.
  • a "wound” is a lesion which results from a bodily injury caused by physical means. Lesions which do not readily heal can be manifestations of conditions, diseases or infections, for example, cutaneous ulcers, osteomyletis, acne vulgaris, draining fistulas, etc. Not uncommonly, lesions do not heal properly and continue to drain which results in discomfort to the patient and a continued threat of severe infection. Such conditions in which tissue does not readily grow to heal the lesion or wound can be the result of bacterial infection or other causes not fully understood.
  • the instant compositions are applied to the affected area or afflicted situs of the patient.
  • topical refers herein to the surface of the epidermal tissue, especially the skin, the surface of tumors on the skin which have been debrided or otherwise modified, as well as sites from which solid tumors have been removed either from the skin or internally.
  • the instant compositions can be particularly useful in conjunction with surgery for removal of internal cancers to eradicate residual tumor cells and act as a prophylactic against local recurrence and metastatic spread of the tumor.
  • the instant compositions can be used instead of surgery when there are cosmetic considerations due to the normally improved appearance of healed situs treated with the instant compositions compared to surgery alone.
  • inj ection can be used for treatment of solid tumors in which removal by surgery is not desired or for which surgery is not medically advisable.
  • the instant composition is injected directly into the tumorous growth.
  • the injection may be accomplished at a number of sites in the growth in order to provide the maximum contact between the instant composition and the tumorous cells.
  • solid tumor refers to tumors in which a plurality of tumor cells are associated with one another, i.e. contiguous and localized within a confined site. This is to be contrasted with "fluid” or “hematogenous” tumors in which the tumor cells occur primarily as unassociated or individual cells , e . g . leukemia. Solid tumors generally propagate on host tissues such as the epithelial, the connective and supportive tissues as well as other tissues located throughout the body.
  • epithelial tumors include papillomas and carcinomas such as squamous cell carcinoma, basal cell carcinoma, adenoma, adenocarcinoma, cystadenoma and cystadenocarcinoma.
  • supportive and connective tissue tumors include sarcomas and their benign counterparts such as fibrosarcoma, fibroma, liposarcoma, lipoma, chondrosarcoma, ⁇ hondroma, leiomyosarcoma and leiomyoma.
  • tissue tumors include gliomas (brain tumors) and malignant melanomas.
  • compositions of the instant invention have been found to be particularly effective against the following solid mammalian tumors: human tumors including malignant melanoma, squamous cell carcinoma, lung squamous cell carcinoma, breast adenocarcinoma, glioma, glioastrocy toma, renal-cell carcinoma, colon, and basal cell epithelioma; canine tumors including mast cell carcinoma, squamous cell carcinoma, mammary adenoma, breast adenocarcinoma, perianal adenocarcinoma, perianal adenoma, sebaceous adenoma, and basal cell carcinoma; and equine tumors including papilloma, malignant melanoma, sarcoid and squamous cell carcinoma.
  • compositions are commonly initially tested by in vitro screening methods. When tested against microorganisms, the composition is commonly applied to a colony at different concentrations and the kill ratio determined. In the treatment of tumors, initial screening is commonly done by the human tumor clonogenic assay. It has been reported that clinical correlations from retrospective analysis and prospective clinical trials with such clonogenic assays have indicated that there is a 60 to 70 percent correlation between in vitro sensitivity and clinical response. The studies have also indicated that there is a greater than 90 percent correspondence between in vitro resistance and treatment failure.
  • the screening of new antitumor agents is still primarily being conducted using a variety of tumor models in vivo.
  • the National Cancer Institute is currently using in vivo tumor models which include the L-1210 lymphocytic leukemia, B-16 melanoma, M-5076 carcinoma, 3 transplantable murine tumors, and the MX-1 human mammary tumor xenograph.
  • the catecholic butane is normally mixed with a suitable solvent.
  • solvents which are effective for this purpose include ethanol, acetone, acetic acid, aqueous alkaline solutions, dimethyl sulfoxide, glycerine, glycerol, propylene glycol, suitably high boiling ethers, nonoxynol, polyethylene glycol, etc.
  • the zinc ions commonly in the form of a toxicologicallyacceptable salt, are mixed with a suitable solvent such as water or polyethylene glycol of low molecular weight, e.g. 200-400.
  • the ionic zinc can be added in the form of readily available salts such as acetates or other aliphatic acid salts while the preferred anion, e.g. chloride, can be added in the form of its readily available salts such as sodium chloride. In the event there is not complete solubilization, the mixture can be milled to obtain a fine suspension.
  • the catecholic butane composition and the source of ionic zinc are mixed in appropriate amounts to achieve the desired concentrations .
  • Additives , adjuvants , other carriers, etc. can be introduced at any stage of the preparation as appropriate.
  • the ordering of mixing of ingredients and the pH of the formulation can be critical.
  • compounds which can serve as counter-ligands are preferably provided so that discreet "molecular" entities are formed rather than polymers of indeterminant length.
  • Such counter-ligands include ethylenediamine tetraacetic acid (EDTA), ethylenediamine diacetic acid (EDDA) , ethylenediamine, ammonia, ethanolamine, amino acids, etc.
  • EDTA ethylenediamine tetraacetic acid
  • EDDA ethylenediamine diacetic acid
  • amino acids etc.
  • Both types of tumors were grown intradermally or subcutaneously in the mice.
  • the B-16 melanoma was grown in BDF. mice and the S-180 tumor was grown in ICR mice.
  • Each mouse was injected intradermally with about 0.01 ml of a saline suspension containing about 1 x 10 6 cells of the tumor cells per 0.01 ml into a preshaven area on the back of the neck of the mouse.
  • the tumors were allowed to grow until they had an approximate size of about 25-100 mg, calculated by the length of the tumor multiplied by the width and height of the tumor measured in millimeters and dividing the product by two.
  • the animals with tumor sizes outside of the size range were culled and the remaining animals were randomly divided into control and test groups.
  • the tumors When the tumors had reached the appropriate size, usually at about day six, the tumors were punctured uniformly,and then treated with either a test compound or a control by topical application to the surface of the tumor. Generally, two topical applications were made 24 hours apart. The materials were applied to obtain from about a 1 to about 2 mm coating over the surface of the tumor. The animals were thereafter observed and their weights and the size of their tumors were periodically measured.
  • the nordihydroguaiaretic acid used in the instant Examples was the meso-isomer and is designated NDGA.
  • Other isomers are indicated, e.g., d,1-NDGA.
  • EXAMPLE 1 A variety of mixtures containing nordihydroguaiaretic acid (NDGA), quercetin, zinc chloride, ascorbic acid or sodium ascorbate were prepared and formulated into a. polyethylene glycol (PEGO) base to obtain an appropriate consistency for application.
  • the mixtures were prepared by dissolving the NDGA in ethanol while warming and stirring. After the NDGA was dissolved the quercetin was added with continued stirring and warming. Upon its dissolution a small amount of water was added and then zinc chloride added or, alternatively, zinc chloride dissolved in water was slowly added to the mixture with continued stirring and warming. Finally, the ascorbic acid or sodium ascorbate previously dissolved in water was then slowly added with continued stirring.
  • NDGA nordihydroguaiaretic acid
  • quercetin zinc chloride
  • ascorbic acid or sodium ascorbate was then slowly added with continued stirring.
  • Example 1 d,1-NDGA used.
  • EXAMPLE 2 The mixtures of Example 1 were tested for antitumor activity against B-16 melanoma and S-180 solid tumor grown in vivo in mice according to the previously described procedure. The results of the tests against B-16 melanoma are given in Table 2 and the T/C tumor size, etc. were determined between day 21 and day 24 post tumor inoculation.
  • Mixtures 21-24 were prepared by dissolving the NDGA in warm (about 65°C) absolute ethanol and dissolving the zinc chloride in the ionized water.
  • the two solutions were then slowly mixed together with stirring and warming until approximately 70% of the solvents were evaporated.
  • the mixtures were then formulated into a PEGO base to obtain a consistency suitable for topical application.
  • Mixtures 25 and 26 were prepared by dissolving the
  • NDGA in a portion of the PEGO by stirring it while warming.
  • the zinc chloride was dissolved in water and warmed while added to the remaining PEGO. The two solutions were then mixed while still warm and stirred until cooled to room temperature.
  • compositions of the various NDGA and zinc chloride mixtures are given in Table 3 (all amounts are given in approximate weight/weight percents).
  • EXAMPLE 4 The NDGA and zinc chloride mixtures of Example 3 were tested for their potential antitumor activity against B-16 and S-180 tumors grown in mice in accordance with the procedure previously described. The results are given in Table 4.
  • EXAMPLE 5 Several different mixtures of butylated hydroxytoluene (BHT), edetic or ethylenediaminetetraacetic acid (EDTA), NDGA and zinc chloride were formulated with a PEGO base. The mixtures were prepared by stirring together and warming a portion of the PEGO and all of the BHT and NDGA until a clear solution was obtained. The zinc chloride, which was weighed quickly to avoid increased weight of absorbed moisture, was dissolved in water with warming. Thereafter, the EDTA was added to the zinc chloride solution with stirring and warming until it was dissolved.
  • BHT butylated hydroxytoluene
  • EDTA ethylenediaminetetraacetic acid
  • the zinc chloride - EDTA solution of mixtures 35, 36, 37 and 38 were titrated with a 50 wt/wt percent solution of sodium hydroxide to obtain pH's of 4.5, 4.5, 2.0 and 3.0, respectively.
  • the remainder of the PEGO 400 was added, with stirring, to the zinc chloride and EDTA until a clear solution vas obtained.
  • the resulting two solutions were then mixed together, while still warm, and stirred until cooled to room temperature.
  • compositions of the mixtures are described in Table 5.
  • 1 PEGO had molecular weight of 200.
  • 2NDGA is d,1-NDGA 3
  • Disodium salt of EDTA 4 NaOH constituted about 17% (wt/wt) of the mixture 5
  • NaOH constituted about 4% (wt/wt) of the mixture
  • 6NaOH constituted about 9.5% (wt/wt) of the mixture
  • NaOH constituted about 1.4% (wt/wt) of the mixture
  • Example 6 The mixtures of Example 5 were tested for their potential antitumor activity against B-16 melanoma grown in mice in accordance with the procedure previously described. The results are given in Table 6.
  • EXAMPLE 7 Mixtures of zinc chloride, EDTA and NDGA were prepared and formulated in a PEGO base. The mixtures were prepared by dissolving the NDGA and EDTA in a portion of the PEGO base by warming and stirring, until dissolved. The zinc chloride was dissolved in water and warmed. The warm zinc chloride solution was added to the warm PEGO containing the NDGA and EDTA and stirred until cooled to room temperature. The composition of the mixtures is given in approximate weight/weight percentage as set forth in Table 7.
  • 1d,1-NDGA was utilized. 2 50 w/w% NaOH was added to ZnCl 2 solution until a pH of about 4.5 was obtained. NaOH equaled about 17 w/w% of final mixture.
  • EXAMPLE 8 The mixtures of Example 7 were tested for their potential antitumor activities against B-16 melanomas grown in mice in accordance with the procedure previously described. The results are given in Table 8.
  • EXAMPLE 9 A mixture comprised of BHT, EDTA, NDGA and zinc iodide was prepared and formulated into a PEGO base. Zinc iodide was dissolved in warm water and EDTA was added with stirring until it was almost completely dissolved and turned yellow. BHT and NDGA were added to warm PEGO and this mixture was then added to the still warm zinc solution. Rapid stirring of the combination of these two mixtures in an ice bath resulted in a thick brown product.
  • composition of the product in approximate w/w% is set forth in Table 9.
  • Example 9 The mixture of Example 9 was tested for its antitumor activity against B-16 melanomas grown in mice in accordance with the procedure previously described. The results are given in Table 10.
  • EXAMPLE 11 Mixtures of zinc chloride and BHT; zinc chloride, EDTA and BHT; zinc chloride and EDTA; zinc chloride, NDGA and BHT; and zinc chloride and ascorbic acid were formulated into PEGO bases.
  • the method of formulation was as follows: anhydrous zinc chloride was weighed quickly to avoid increased weight of absorbed moisture, placed into a beaker with water warmed in a water bath at 70-80°C and stirred until dissolved. If EDTA was a component, it was added with stirring to the warm zinc mixture until dissolved. When BHT or NDGA were included in a mixture, either or both were added to PEGO warmed in a water bath at 70-80°C and stirred until a clear solution was obtained. The two solutions were then mixed while still warm and stirred until cooled to room temperature and then refrigerated.
  • the zinc chloride, ascorbic acid solution was prepared by dissolving a measured quantity of each component in water, combining the two mixtures, then adding ethyl alcohol and PEGO.
  • the compositions of the mixtures are given in Table 11.
  • EXAMPLE 12 The mixtures of Example 11 were tested for their potential antitumor activity against B-16 melanoma grown in mice in accordance with the procedure previously described. The results are given in Table 12.
  • compositions of the mixtures as weight percent of the total composition are given in Table 13.
  • Mixtures 62, 63 and 68 were prepared according to the following method. NDGA was dissolved with stirring in warm (65°C) ethanol and quercetin was added with stirring until the solution was clear. Deionized water in which the sodium ascorbate had been dissolved was added dropwise with stirring. Then the metal salt was added and the stirring continued until the salt was dissolved. The solution was evaporated to dryness in a vacuum and these stored in an air-tight vial until mixed with the necessary amount of PEGO.
  • Mixture 64 was prepared by dissolving the NDGA in a minimal amount of ethanol. The cadmium salt and ascorbate were dissolved in deionized water and added to the NDGA solution dropwise with stirring and warmed to
  • the mixture was evaporated to about 130% dry weight and formulated with the PEGO base.
  • mixture 65 the salt was dissolved in water and mixed with PEGO base.
  • Mixture 66 was prepared by dissolving the NDGA, ZnCl 2 and CuCl 2 2H 2 O in ethanol. The copper caused oxidation of NDGA and a precipitate. The ethanol was evaporated, and PEGO was added to the product. Mixture 67 was prepared in the same manner as 66, except that the cadmium salt was added in ethanol and water. A precipitate formed on evaporation and was mixed with PEGO. The product changed in consistency depending upon the temperature.
  • Example 13 The mixtures of Example 13 were tested for their potential antitumor activity against Sarcoma-180 tumors grown in mice in accordance with the procedures previously described. Test results are given in Table 14.
  • EXAMPLE 15 Six different mixtures were prepared having the compositions shown in Table 15.
  • Mixtures A-E were prepared by mixing the quercetin, NDGA and sodium hydroxide crystals together in approximately 300-400 milliliters of water. The mixture was heated to approximately 60°C and it took approximately 1/2 to 1 hour for the mixture to dissolve while being stirred constantly. The resulting solution had a red-orange color.
  • zinc chloride and/or sodium ascorbate were added, they were first dissolved in approximately 300 milliliters of water. Then the two solutions were mixed together with stirring to form the mixtures. Immediately upon their intermixing a cottage cheese mixture was formed which, with continued stirring, became a slurry. The slurry was then roto-evaporated to dryness.
  • Mixture F was prepared by putting the zinc chloride into solution in water and then adding sodium ascorbate to zinc chloride solution.
  • the NDGA was added to the zinc chloride and sodium ascorbate aqueous solution. The solution was a little cloudy indicating that not all of the NDGA was in solution.
  • Approximately 10 drops of concentrated sodium hydroxide (approximately 40%) were added and the solution cleared a little .
  • the quercetin was then added s lowly with constant stirring and another approximately 5 drops of concentrated sodium hydroxide was added dropwise. The solution turned yellow to slightly orange and a flocculant precipitate formed. The precipitate was then dried by evaporation in a rotovap at approximately 50-60°C.
  • Each of the mixtures was formulated into a PEGO (polyethylene glycol) base.
  • Example 15 The mixtures of Example 15, in addition to controls, were tested against xenografts of the transplanted human lung squamous cell carcinoma, LX-1, and human breast adenocarcinoma, MX-1 in athymic (nude) mice of BALB/c background. Each animal was inoculated intradermally on the dorsum near the nape of the neck with 0.05 ml. of a LX-1 or MX-1 tumor homogenate. Tumor weights, in milligrams, were calculated from the measurement of the length (L), width (W) and height (H), in millimeters of the tumors using the formula (LxWxH)/2. The animals were randomized in groups to ensure representation of smaller and larger tumors.
  • Topical treatment of the tumors was utilized and to assure penetration of the mixtures, the tumors were punctured with an 18 gauge (1-1/2 inch) needle to a depth slightly above the bevel (3/16 inch).
  • the number of punctures varied from 8 to 12 depending on the size of the tumor.
  • Experiment 1 the animals were treated twice, 18 and 19 days after LX-1 tumor inoculation.
  • the animals of Experiment 2 were treated once at day 25 after LX-1 tumor inoculation.
  • the animals of Experiment 3 were treated once at day 23 after MX-1 tumor inoculation.
  • the results of Experiments 1, 2 and 3 are given below in Tables 16-A, 16-B and 16-C.
  • the mean delta tumor weight is the difference in mean tumor weight between the day specified and the mean tumor weight on the day of treatment.
  • the tumor growth inhibition (positive or zero mean delta tumor weight) is expressed as a %T/C value calculated from the average test delta TW/average control delta TW and may be positive or zero.
  • Tumor regression (negative delta TW) is expressed directly as a percentage of the test change in tumor weight to the initial tumor weight and is preceded by an R. For each experiment the groups of mice treated with mixtures A-F are compared with the group of mice treated with the PEGO control.
  • EXAMPLE 17 To 36.7 grams of powdered Larrea divaricata were added 24.5 grams of powdered rosehips and the mixture was blended in a blender for 5 minutes. The blended mixture was then mixed with 100 milliliters of an aqueous solution containing 185.9 grams zinc chloride to form a paste. The paste was allowed to stand at room temperature for 24 hours. Thereafter, it was stirred and then placed in a screw-capped glass container. The container was placed in a humidified oven at 40°C for 5 days. This incubated paste was then suspended in 500 milliliters of triple distilled water and shaken at room temperature for 24 hours on a reciprocating shaker.
  • the zinc chloride extract solution was then evaporated to near dryness on a rotary evaporator at 90°C under reduced pressure. A sufficient quantity of this dried zincchloride extract was added to 120 grams of an ointment base consisting of 10 % (w/w) stearyl alcohol and 90%
  • a sufficient quanitity of the paste of Example 17 was added to sterile deionized water to obtain a concentration of 10 grams per 100 milliliters of water.
  • the aqueous mixture was thoroughly shaken for one hour on a reciprocating shaker, then the aqueous suspension was filtered through Whatman #1 filter paper in a Buchner funnel. The filtrate, an aqueous suspension, was used to irrigate wounds in the treatment of osteomyelitis.
  • Example 18 Five selected human patients with osteomyelitis of duration of from several months to several years were treated topically with the solution of Example 18 and/or the paste of Example 17. In all instances, the osteomy elitis had been unresponsive to conventional treatment, and upon the application of the preparation, the patients received no other conventional therapy except as indicated. In some cases, the wounds were debrided, prior to the application of the preparation. Upon application of the preparation, most patients experienced pain and a burning sensation over the area which had been treated and some patients additionally experienced swelling and inflamation. One patient experienced severe nausea after an application of the preparation.
  • EXAMPLE 20 Fifteen older dogs having perianal adenomas were treated topically with the ointment of Example 17 having a strength of 55% (w/w). The normal treatment for such a condition is surgery; however, these older dogs were poor surgical risks. The tumor of each dog was biopsied and the ointment was applied topically into the biopsied incision. The duration of treatment varied depending upon the severity of the adenoma. Dogs with simple circumscribed adenomas required only one treatment. The dogs with more advanced adenomas generally required more than one treatment which were given three to five days apart. The treatment was successful in thirteen of the fifteen dogs. The treatment was not successful in two of the dogs which had extremely advanced cases of perianal adenomas.
  • EXAMPLE 21 An incubated paste of rosehips, zinc chloride and Larrea divaricata prepared in accordance with the method of Example 17 was placed into gelatin capsules such that each capsule contained 200 mg of the paste.
  • a patient with glioastrocytoma was treated orally with these capsules. Prior to this treatment the patient had a resistant tumor which displaced the cranium and protruded from the right lateral aspect of the skull; the protrusion measured 7 x 7 mm.
  • the patient received 200 mg oral doses four times a day for a total daily dose of 800 mg. Observable and subjective improvement occurred within seven days; in 71 days the tumor had become cystic and lysed.
  • the protuberance of the skull was reduced to near normal dimensions by repeated aspirations of the clear amber cystic tumor fluid.
  • the patient has been maintained on the 200 mg capsules given four times daily and has remained symptom free for over 18 months.
  • Examples 22 through 35 describe the results of an investigation into the antineoplastic activity of a number of compositions in a series of experiments against human cancer xenografts implanted in athymic Balb/C (nude) mice.
  • the mice were maintained under special research conditions which included positive laminar flow ventilation and sterilized food and water. Animals were identified with standard ear tags.
  • mice When the mice were six to eight weeks old, a fragment, weighing approximately 25 mg., of a human cancer was implanted subcutaneously in the left flank under anesthesia and the incision was closed and allowed to heal. The tumors were allowed to grow until they reached an approximate size of between 25 and 100 mm 2 (length x width). The approximate weight of the tumor was determined by measurement with vernier calipers, according to the formula:
  • a x B x C Weight (mg.) 2
  • A length in mm.
  • B width in mm.
  • the human tumors investigated were breast adenocarcinoma - MX-1, lung squamous cell carcinoma - LX-1, renal cell cancer, a brain cancer (glioma), a melanoma, and a colon cancer.
  • mice Five mice were treated with the test composition for most experiments. The mice were anesthetized and given a single 0.05 ml. or 0.10 ml. intratumor injection of the test composition using a 23 or 27 gauge needle.
  • the test composition administered in the following Examples is
  • necrotic or cratered skin tissue resulted. The size of this necrotic area was measured as length times width and the height was recorded as zero. After the cratered area healed somewhat, it was possible to palpate the area and note whether a tumor remained, or regrew.
  • mice were observed for a total of approximately 60 days, and were then sacrificed.
  • the following categories of animal response were decided upon to summarize and report treatment results.
  • Tumor Free at 60 Days - indicates an animal that has survived the entire 60 day test period and is free of tumor at the end of that time.
  • Tumor at Death the animal has a tumor when it dies or is sacrificed. The end point may occur before the 60 day period is complete.
  • Premature Death the animal fails to survive the full 60 days having died of unknown causes, or is sacrificed when obviously ill to avoid cannibalization.
  • the category does not include animals sacrificed due to massive tumor growth, those animals dying of anesthetic overdose or tho which are actually cannibalized by their cage mat
  • Tumor Recurrence - includes animals in which the tumor reappeared after an earlier tumor-free period. These are animals in which the original eschar has resolved, and palpation has indicated an absence of a tumor for some period; however the tumor has now regrown. Animals may be reported in more than one category. For example, an animal found dead in its cage but having a tumor would appear in both the "Premature Death" and "Tumor at Death” categories.
  • Test compositions were prepared according to the following general method.
  • a stock solution of zinc chloride was prepared by dissolving a measured amount in water with heating and stirring - NDGA, or its analog desmethyl NDGA, was dissolved in PEGO 400 with heating and stirring. The two components were mixed together and allowed to cool with vigorous stirring. Additional PEGO 400 was added to further dilute the test compositions to achieve the approximate concentrations in wt/wt % given below. More than one stock solution was prepared according to the above-described procedure; the various test compositions were tested at different times.
  • test mixtures used were prepared by the following general method. In each case, any missing component was replaced by an equal amount of PEGO 400. If the general method differs, that difference is noted in the specific example.
  • NDGA is included in a mixture, it is dissolved in PEGO
  • BHT butylated hydroxytoluene
  • EDTA ethylenediaminetetraacetic acid
  • Control - polyethylene glycol 400 Additional mixtures of varying concentrations of zinc chloride, NDGA and EDTA were also prepared as described above and the pH of the mixture was measured. All of the mixtures were tested against human breast adenocarcinoma, MX-1, grown in five athymic Balb/c mice following the procedure previously described.
  • test compositions were prepared according to the general method described in Example 23 except that the composition of Group C was treated in the following manner. After the zinc chloride and EDTA were dissolved in the warm water, this test solution was titrated with a 50% solution of NaOH in water until the zinc solution reached pH 4.5. In all other respects preparation remained the same.
  • compositions were tested for antitumor activity in five athymic Balb/c mice implanted with human breast adenocarcinoma, MX-l, according to the protocol previously described. Test results are given in Table 24.
  • EXAMPLE 25 The following compositions were prepared according to the general method described in Example 23. The few changes in method are described after listing of test compositions. Compositions of test materials were as follows:
  • Control Polyethylene glycol (PEGO 400)
  • the mixtures of Groups A through E each had the following components in common:
  • Maracarb and Kelig-32 are trademarks for wood-derived products of American Can Company, Lignin Chemicals, American Lane, Greenwich, Connecticut 06830.
  • the compositions are soluble in water and able to chelate metal ions.
  • the lignins, Marcarb and Kelig-32 were dissolved in cold water to which was then added the ZnCl 2 .
  • the EDTA was the third component added to the aqueous solution and was warmed with stirring until dissolved, or in the case of the Kelig-32 solution, until the EDTA was almost dissolved. The rest of the general procedure was followed, but the resulting product was then milled in an ink mill at 0.002".
  • the mixtures were tested for antitumor activity in five Balb/c mice implanted with human breast adenocarcinoma, MX-1, following the protocol hereinbefore described.
  • EXAMPLE 26 Test compositions were prepared according to the following general method.
  • the NDGA, BHT, and PEGO 400 were measured and mixed together with heating until melted and dissolved.
  • the PEGO Base consisting of 50% Pego 400, 45% Pego 3350 and 5% stearyl alcohol, was prepared by mixing and heating the components together in a separate container until they dissolved.
  • ZnCl 2 and EDTA were dissolved in water with heating and stirring in a separate container.
  • the ingredients in each of the separate containers were added together in amounts needed to give the concentrations desired and allowed to cool with vigorous mixing. Any further dilution to achieve desired wt/wt % was achieved by adding Pego 400. When an ingredient was omitted from a particular composition, the amount of the missing ingredient was supplied by adding additional Pego 400.
  • test compositions were tested in five athymic mice implanted with human breast adenocarcinoma, MX-1. Results are given in Table 26.
  • compositions Two test compositions were prepared according to the general procedure previously described. However, in Composition 1 the zinc chloride was replaced by zinc iodide, and in Composition 2 the zinc chloride was replaced by zinc bromide. Approximate concentrations of the ingredients are given below in wt/wt percent.
  • Test compositions containing a proprietary slowrelease formula of KV-Pharmaceutical Co. and varying proportions of BHT, EDTA, NDGA and zinc chloride was prepared according to the general method described in Example 26.
  • the proportions of BHT, EDTA, NDGA, and zinc chloride are shown in Table 28.
  • the remaining ingredient in the test composition and the control comprised the proprietary compound.
  • test composition was investigated for its antineoplastic activity against xenografts of the following human cancers: lung squamous cell carcinoma, LX-1; breast adenocarcinoma, MX-1; renal cell cancer; brain cancer (glioma); melanoma; and colon cancer.
  • the test composition with the approximate wt/wt percentages given below was prepared according to the procedure previously described in Example 26.
  • a control composition consisting of Pego 400 was also prepared.
  • Pego 400 88.77 100 The composition was then tested for its effect on human tumors of varying origin implanted in athymic mice as previously described. Generally, there were ten mice in each group tested with composition 1, and five mice in each group tested with Pego 400 control. Instances in which the number of mice varied are specifically indicated.
  • mice tested One mouse died from anesthesia overdose the first day of test and was replaced with another mouse.
  • EXAMPLE 30 For Examples 30 through 32, a number of organic compounds were formulated into test compositions according to the following general method. Zinc chloride was dissolved in Pego 400 to prepare a stock solution. The amount of organic compound required to give the final concentration given below was measured into a clean vial and Pego 400 was added with mixing until dissolved.
  • test compositions reported in Table 30 are identified in the table by the organic compound they include, and were tested for their effectiveness as antitumor agents against xenografts of the human breast adenocarcinoma, MX-1, grown in athymic mice, by intratumor injection according to the protocol previously described.
  • EXAMPLE 31 A number of organic compounds were formulated and tesred as set forth in Example 30 with 0.69% zinc chloride, and without zinc chloride. As part of this comparative testing, some of the organic compounds included known anticancer agents. The results of the organic compounds with and without zinc chloride are set forth in Table 31.
  • EXAMPLE 32 A number of organic compounds were tested in varying concentrations for their antineoplastic effectiveness against xenografts of human breast adenocarcinoma , MX-1, and grown in groups of five athymic mice, according to the protocol previously described. All compounds contained 0.69% Zn. Results are set forth in Table 32.
  • EXAMPLE 33 A solution of 4.6% NDGA in polyethylene glycol 400 was diluted 1:20 and 1:80 with polyethylene glycol 400, and injected intraperitoneally into nude mice, bearing transplanted human breast adenocarcinoma, MX-1, tumors. Four mice were injected with the 1:20 dilution, and three mice were injected with the 1:80 solution. All mice were sacrificed on day 19 due to large tumor size. No retardation of tumor growth was observed.
  • compositions were tested to determine the compositions' antineoplastic effectiveness against xenografts of human breast adenocarcinoma, MX-1, grown in groups of five athymic mice, according to the protocol previously described.
  • concentration of the various metal salts in the test compositions was 0.73%
  • EXAMPLE 35 For each composition as set forth in Table 35, two mice without tumors were injected subcutaneously on the flank. Also, for each of the same compositions two mice, each having a transplanted human breast adenocarcinoma, MX-1 tumor, were injected subcutaneously on the flank opposite that bearing the tumor. The injection sites healed well without ulceration or scarring in all cases. The animals with tumors were sacrificed on day 22 due to large tumor size. No retardation of tumor growth was observed.
  • compositions suitable for topical application were prepared, containing zinc chloride (ZnCl 2 ), nordihydroguaiaretic acid (NDGA) , edetic or ethylenediaminetetraacetic acid (EDTA), butylated hydroxytoluene (BHT), stearyl alcohol, purified water, polyethylene glycol having an average molecular weight of 400 (PEGO 400), and polyethylene glycol having an average molecular weight of 3350 (PEGO 3350).
  • the compositions were prepared in the following manner: the purified water was placed in a clean glass container of suitable capacity, the water was heated to about 80-90°C with stirring, and zinc chloride was added to the heated water, continuing the stirring until the zinc chloride dissolved.
  • the edetic acid was next slowly added with mixing until dissolved.
  • the polyethylene glycol 400 was heated to about 80-90°C with stirring, the NDGA was added thereto, then the BHT, and this mixture was added to the zinc chloride-edetic acid solution with stirring. The entire mixture was then cooled to about room temperature and passed through a number 3 roller mill until smooth.
  • the polyethylene glycol 3350 was then heated to about 80-90°C in a suitable container and the milled ingredients added thereto with mixing.
  • the four compositions given below were prepared by the method described above and the final compositions in wt/wt % were as follows: Compound
  • test compositions were tested in clinical studies for its antineoplastic activity on various cancers and diseases on the patients.
  • the protocol for administering the compositions is as follows: for Compound A, no pre-treatment or preparation of the lesion or surrounding skin was done prior to application of the compound. Prior to topically applying Compounds B, C, or D on basal cell epithelioma and actinic keratosis, the surface of the lesions were tape stripped prior to each application. Tape stripping involves pressing a sticky medical or surgical tape on the lesion site, then removing the tape. The test compound was then applied to the lesion. Following the application of the compound, some of the lesions were covered with a dermatological dressing. The test compound remained on the lesion for at least 48-72 hours. A second application of the compound was made 48-72 hours following the first application using the identical technique described above.
  • test compound A Fifty-seven patients with basal cell epithelioma were treated with Compound A, B, C or D.
  • the test medication was applied directly to the lesion with a coating of approximately 2 mm thick and confined to the visual margins of the lesion.
  • the lesion was then covered with a dressing and the patient was advised against washing the treated area for a reasonable period of time as determined by the investigator.
  • a visual examination and measurement of the lesion was performed at 3-4 day intervals.
  • a second application of the same test compound was applied after a minimum of seven (7) days following the initial treatment.
  • an excisional biopsy was obtained 30 days after the initial treatment.
  • Compound A was used to treat eight human patients with basal cell epithelioma. No pretreatments or preparations of the lesion or surrounding skin were done prior to application of the composition.
  • T.W., H.S., and J.H. were treated with a single topical application of the composition.
  • patient J.H. forty days following the single application of the composition, the wound remained crusty and the biopsy was positive for basal cell epithelioma.
  • EXAMPLE 37 Fifty-nine (59) human patients with actinic keratosis were treated with Compound B, C, or D. The test medication was applied directly to the lesion with a coating of approximately 2 mm and confined to the lesion margin. A dressing was applied to the lesion and the patient was advised against washing the treated area for a reasonable period of time. A visual examination and measurement of the lesion was performed 7 and 14 days following the initial treatment. At the discretion of the investigator, a second treatment with the same test compound was applied. In order to determine whether the test compound eradicated the premalignant neoplasm, a punch biopsy was obtained 30-60 days after the initial treatment.
  • Two human patients with recurrent cutaneous chest wall adenocarcinoma of the breast were treated with Compound A.
  • the compound was applied directly on the lesion with a thickness of approximately 2 mm and confined to the visual margins of the lesion.
  • One (1) lesion was punctured with a needle to facilitate penetration.
  • a dressing was applied to cover the treated lesion and the patient advised against washing the treated area for a reasonable period of time.
  • a crust formed at each site of the test compound application and the area of the lesion was well demarcated.
  • a second application of Compound A was applied to three (3) lesions.
  • a biopsy was obtained to determine the effect of Compound A on the malignant neoplasm. If the tumor was completely eradicated, the patient was examined periodically for 12 months. If the biopsy continued to show evidence of recurrent cutaneous chest wall adenocarcinoma of the breast or if the lesion was not clinically improved by the 14th day after the initial treatment, the patient was withdrawn from the study.
  • EXAMPLE 39 Eleven cutaneous ulcers in eight human patients were treated with Compound C. If excessive necrotic material was present, debridement of non-viable and foreign material was performed either surgically or with wet-dry dressings prior to treatment. The test compound was applied directly to the cutaneous ulcer in an amount sufficient to cover the visual margins of the ulcer. The treated ulcer was then covered with a loose dressing and the patient advised against washing the treated area for a reasonable period of time. A scab or crust was observed to form on the surface of the ulcer. Normallv within two weeks the crust had loosened to where it was sluffed off or could be readily removed. It was observed that granulation of the tissue in the ulcer had occurred in those ulcers which showed clinical improvement.
  • a second treatment with Compound C was applied after removal of the crust.
  • the patient was visually examined and the ulcer measured within two weeks after the initial treatment. Thereafter, the patient returned twice a month for two months for a visual examination and measurement of the ulcer.
  • eleven (11) treated lesions seven (7) were clinically improved.
  • EXAMPLE 40 Six (6) Kaposi's sarcomas in human patients were treated with Compound A, which was applied directly to the lesion with a thickness of approximately 2 mm and confined to the visual margins of the lesion. The lesion was then covered with a dressing and the patient advised against washing the treated area for a reasonable period of time. The patient was visually examined 1, 2, 3, 7 and 14 days after the initial treatment. If possible, accurate measurements of the lesion were taken and recorded. A second application of Compound A was applied as deemed necessary. After 14 days, a biopsy was obtained if the lesion appeared clinically improved.
  • EXAMPLE 42 Canine patients were treated with Compounds A, C, D or E according to the protocol of Example 41. The results are given in Table 42.
  • Equine patients with various tumor lesions were treated with Compound A, C or D.
  • Melanoma, sarcoid and squamous cell carcinoma lesions were removed to skin level by surgical debulking; for papillomas, the lesion tips were removed.
  • the tumor site was covered liberally with the test compound extending 5 mm peripherially.
  • the crust was removed, the lesion area abraded and the test compound applied topically. After an additional two weeks, any crust was again removed from the lesion and the area abraded. The same test compound was again applied topically.
  • a biopsy of the lesion area was performed. The results of the equine studies are shown in Tables 43A and 43B.
  • EXAMPLE 44 Equine patients were treated with Compounds A, C, D or E according to the protocol of Example 43. The results are given in Table 44.
  • the 14 C-NDGA compound exhibited a specific radioactivity of 20.2 Ci/mol (66.9 micro Ci/mg) and a purity of 96.9% by mass spectrometry and by radioautography of thin-layer chromatography plates developed in benzene: isopropanol: acetic acid:water (25:5:2:10).
  • Compound A showed the final mixture to be homogeneous with regard to 14 C and NDGA; it contained 53 micro g of
  • the compounds were dermally administered to young adult Sprague-Dawley rats by the following protocol:
  • the back skin of the rat was prepared by removing the hair from a 5 x 5-cm area with a clipper and the residual hair stubble was removed with a wax expilatory. Then the skin was stripped repeatedly (5x) with adhesive tape until the stratum corneum was removed. Then 0.5 gm of the formulation was weighed on a
  • Compound A devoid of Zn and EDTA was performed on 15 male Sprague-Dawley rats (mean weight 241 ⁇ 7 g). They received an average of 0.390 ( ⁇ 0.019) g of Zn-free C205 containing 14 C-NDGA. The average dose of 1 4 C-NDGA was 83.2 mg/kg of body weight. Groups of three rats were bled terminally and tissues were taken at 4,
  • Table 45D lists the compositions of the compounds and the amounts of materials used for preparing the compounds containing 14 C-NDGA. These compounds were analyzed for
  • the mean rat body weights, average doses of. the formulations, and mean doses of 14 C-NDGA in mg/kg of body weight for the three current protocols were: 297 ⁇ 15 g (standard deviation), 512 + 28 mg, and 82.7 ⁇ 2.0 mg/kg for Compound D; 325 ⁇ 12 g, 570 ⁇ 26 mg, and 84.0 ⁇ 1.4 mg/kg for Zn-free Compound D; and 328 ⁇ 27 g, 575 ⁇ 45 mg, and 84.2 ⁇ 2.8 mg/kg for modified Compound D.
  • One milliliter of the bacterial, yeast, or mold spore suspension was transferred to a sterile 12-ml glass, conical, centrifuge tube covered with a sterile cap and centrifuged at 3,000 rpm for 15 min. Centrifugation was done at room temperature using a benchtop, angle-head, clinical centrifuge (Clay-Adams). After the bacteria, yeast, or mold spores were pelleted, the supernatant fluid was decanted and the tubes inverted over paper saturated with a biocide placed in a bacteriological hood.
  • the pellets in the centrifuge tubes were then mixed with 1 gram of the undiluted test material and allowed to remain in contact for 2 hours at 37°C for the bacteria and yeasts and at 25°C for the mold spores.
  • the test mixture was diluted 1 to 10 with growth medium (TSB or SAB broth). Additional serial dilutions were made from the initial dilution up to 1 x 10 -9 . Each material was tested in triplicate.
  • the controls which consisted of the microbial cells incubated with 1 gram of mineral oil, were diluted in the same way. All dilutions of both test materials and controls were then incubated at an appropriate temperature of 37°C for bacteria and yeasts and 25°C for molds to allow for growth of any viable cells present.
  • M. smegmatis All bacterial species except M. smegmatis were incubated for 48 hours; M. smegmatis was incubated for 7 days. Yeast tests were incubated 48 hours. Molds were incubated for 10 days. For a determination of growth response, growth in tubes containing test compositions was compared to the growth in a mineral oil control at an equivalent dilution. Growth was indicated by turbidity in the broth medium.
  • EXAMPLE 47 Tests were conducted on the effect of the direct exposure of representative microorganisms to several separate components of the composition of Example 36. Pego base alone was tested in one series of evaluations to determine whether or not inhibition by this carrier would have to be considered in evaluating the results of the individual ingredients dissolved in it.
  • the amount of polyethylene glycol present in the formulation was calculated.
  • the pure base material was then diluted with water to this concentration.
  • Mineral oil was used as a positive control.
  • Nordihydroguaiaretic acid (NDGA) and desmethyl NDGA (DM-NDGA) diluted in pego base were also tested for inhibitory properties against representative gram-negative and gram-positive bacteria and yeasts covering the spectrum of microorganisms used in these tests.
  • the initial concentration of the compounds tested was equivalent to the amount present in the composition, and the general procedure outlined in Example 46 was followed. After a 2-hour exposure of the microorganisms to this initial concentration, progressive 1 to 10 serial dilutions of the mixture were made to assess viability of any microorganisms present. Results are shown in Table 47.
  • EXAMPLE 48 A combination of EDTA (ethylenediaminetetraacetic acid) and zinc chloride in pego base at the concentration in which these components are present in composition A of Example 36 was tested for its effect on the viability of representative microorganisms. All organisms given in Example 46 were tested except Candida Kruseri, and
  • Example 46 The test procedure followed was that generally described in Example 46.
  • EXAMPLE 49 A test was conducted to assess the growth of Escherichia coli and Staphylococcus aureus in broth containing composition A of Example 36, NDGA or desmethyl NDGA diluted in glycerol. Test parameters and results are given below.
  • EXAMPLE 50 A series of broth dilution tests were conducted to assess the effect of composition A of Example 36 and its separate components on the growth of microorganisms.
  • the individual test materials were incorporated into pego base at the concentration in which they are present in the composition for testing.
  • EDTA and zinc chloride were tested together.
  • Each original formulation was diluted 1 to 10 with growth medium, (usually tryptic soy broth with glucose), and subsequent 1 to 10 dilutions were made of the previous dilution usually up to 1 x 10 -4 . This test was done with no consideration given to the solubility of the test material when diluted. In all cases, controls consisting of cells in mineral oil diluted in TSB were made to test the effect of the medium on growth.
  • each series of materials was made by testing a duplicate set of tubes that were uninoculated.
  • Each dilution tube containing 10 ml. test broth was inoculated with 0.1 ml. of a 24-hour culture of all test species except M. smegmatis and the mold species.
  • Spore suspensions of fungi (10 days) were used to inoculate the tubes for testing effects on M. canis and T. mentagrophytes, and Sabouraud's broth was used for dilution because the fungal species grew somewhat better in this medium than in TSB.
  • a stationary phase culture of each test species was used.
  • Results of the broth dilution tests are given in Table 50.
  • the pH values given in the tables apply only to the dilution shown.
  • a 4+ reading for one microbial culture does not mean that the turbidity of that culture was the same as a 4+ reading for any other culture.
  • a 4+ reading means that turbidity in the tubes of a particular test was equal to the turbidity of the appropriate control at the dilution compared.
  • EXAMPLE 51 Compositions containing NDGA, zinc chloride or a combination of NDGA-Zn were tested for antimicrobial activity individually against seven gram positive and gram negative bacterias, yeasts and molds.
  • a vehicle containing 30% polyethylene glycol-200 (PEGO-200) + 0.1% sodium ascorbate in deionized water at various concentrations was shown to exhibit no inhibitory effect on microbial growth and was chosen as the diluent for the test compounds.
  • Stock solutions of the test compounds in 30% PEGO-200/water were prepared at the following weight percent concentrations: 4.6% NDGA + 0.1% ascorbic acid; 5.0% ZnCl 2 + 0.1% sodium ascorbate; and 4.6% NDGA + 5.0% ZnCl 2 + 0.1% sodium ascorbate. Aliquots of the stock solutions were diluted 1:10 and 1:100 with the 30% PEGO-200 diluent.
  • the stock solutions were further diluted 1:10 with Brain Heart Infusion Agar, which was melted at 45°C prior to the addition of the test solutions.
  • the agar containing the test solutions was then poured into 50 x 90 mm petri dishes and allowed to dry for four hours at room temperature prior to inoculation.
  • All Brain Heart Infusion slants were started at 35°C anaerobically except for T . mentagrophytes at 27°C and P. acnes at 35°C anaerobically. Those microbial slants incubated at 35°C were subsequently transferred to new slants at 35°C and incubated at the same temperature. All slants were harvested with 1 ml saline containing 0.05% Tween-80 and diluted with saline in the following amounts to be used as working inocula: 1 ml each of E. Coli, P. aeruginosa, S. aureus and B. subtilis was diluted with 99 ml saline; 1 ml of C. albicans and P.
  • the in vitro antitumor effect of the interaction of NDGA and ZnCl 2 at various ratios were determined utilizing the clonogenetic assay.
  • the human lung tumor cell line, LX-T which was derived from the solid tumor, LX-1, was cultured in the standard medium RPMI-1640 + 10% fetal calf serum (FCS).
  • a stock solution (10 -2 M) of NDGA was prepared by dissolving 32.04 mg of NDGA in 4 ml DMSO and 6 ml distilled H 2 O.
  • a 10 -2 M stock solution of ZnCl 2 was prepared by dissolving 13.63 mg ZnCl 2 in 10 ml of distilled H 2 O.
  • HBSS Hank's balanced salt solution
  • Freshly prepared single cell suspensions of LX-T cells were analyzed by foward and right angle light scatter using the Ortho System 50 Cytofluorograf with the coherent 5-watt Argon ion Laser and 2150 computer system. An integration boundary was formed around the single and double cell light scatter distribution. A second boundary was established which included the single cells and colonies. These boundaries excluded any small particles which were carried over from the agarose washings (1-2 micrometers, um). The diameter range for the LX-T cells was 15-20 um, while the colonies formed after 3 days incubation were 50-100 um.
  • DNA Flow Cytometric Analysis After LX-T cells were incubated in the presence of the drugs, they were harvested by slight scraping with a rubber policeman ad centrifuged at 500 xg, 100 min., at room temperature. The cells and colonies were resuspended in a solution of the DNA-specific fluorochrome, (4', 6-Diamidino - 2 - phenylindole (DAPI) (polysciences) dissolved in a speical nuclear isolation medium. The suspension of nuclei was passed through a 70 um filter and stored on ice.
  • DAPI 6-Diamidino - 2 - phenylindole
  • Trout red blood cells were used as the DNA standard (5.0 pg/nucleus) for these studies as described previously to quantitate the DNA content of the LX-T nuclei.
  • the DNA distributions of the DAPI-stained nuclei were obtained as previously described.
  • Tables 52A, 52B and 52C contain results using ratios of NDGA:ZnCl 2 of 1:0.5, 1:2 and 1:5 respectively.
  • Tables 52D and 52E contain results using
  • Table 52H contains results of evaluations of three known anticancer drugs.
  • the ED (50) for each was calculated using the data in Table 52H. These values along with ED (50) for NDGA are given in Table 52l.
  • NDGA and ZnCl 2 at various ratios was determined against MX-l (human breast adenocarcinoma) cells.
  • MX-l cells were cultured in the standard RPMI-1640 media and implanted subcutaneously in the flank of nude mice in order to propogate the tumor line. Nude mice were implanted with
  • the effective doses (EDx ) at different response levels were determined mathematically. These values for ZnCl 2 alone, and of ZnCl 2 in combination with NDGA are provided in Table 53E. The effective doses of NDGA alone and of NDGA in combination with ZnCl 2 are provided in Table 53E.
  • Liquid samples were prepared containing 50 volume percent each of water and ethanol, and 10 weight percent NDGA and 2.5 weight percent of the indicated metal ion. The pH of the water was adjusted to either 4, 7 or 10, as indicated.
  • the solutions were placed in quartz tubes in the cavity holder of a Bruker ER 200D electron spin resonance (ESR) spectrometer. Initiation was accomplished with the addition of 1 weight percent of ammonium persulfate and 0.05 weight percent N,N,N',N'-tetramethylenediamine (TMD) dissolved in ethanol.
  • ESR electron spin resonance
  • Table 54C contains values for relative ESR signal intensity at different time periods after initiation at a solution pH of 10 for NDGA alone, NDGA with zinc ions, and NDGA with magnesium ions.
  • NDGA was separately tested with ferrous ions and cobalt (II) ions at a solution pH of 7. No ESR signal could be detected.
  • Dopa 3-hydroxytyrosine, was used instead of NDGA both with and without zinc chloride at a pH of 7. No ESR signal was detected with dopa alone.
  • the "slope of curve” is the rate of change of free radical concentration per unit of time determined, from the data in the Tables in the usual way of 10 -8 M.
  • the various rate constants Kd are given in Table 54E.

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Abstract

Sont décrits des compositions pharmacologiquement actives de butanes catécholiques et de zinc ionique et leur utilisation dans le traitement de maladies et d'affections de la peau ainsi que leur utilisation en tant qu'agent anti-bactérien et anti-micosique. Lesdites compositions sont également utiles dans le traitement de tumeurs solides bénignes, prémalignes et malignes, notamment de la peau.
PCT/US1986/001740 1986-08-25 1986-08-25 Compositions pharmacologiquement actives de butanes catecholiqueset de zinc WO1988001509A1 (fr)

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Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0288534A4 (fr) * 1986-10-28 1989-02-23 Chemex Pharmaceuticals Inc Compositions de butanes catecholiques avec du zinc.
EP0290442A4 (fr) * 1986-11-19 1990-07-03 Chemex Pharmaceuticals Inc Composes pharmacologiquement actifs et leurs melanges, compositions organiques et sels metalliques.
WO1990015603A1 (fr) * 1989-06-12 1990-12-27 Shiseido Company, Ltd. Composition antipruritique
US5219847A (en) * 1989-06-12 1993-06-15 Shiseido Company, Ltd. Antipruritic composition
WO1995005156A1 (fr) * 1993-08-17 1995-02-23 Schering-Plough Healthcare Products, Inc. Compositions servant au traitement des cors, callosites et verrues
WO1999017761A1 (fr) * 1997-10-06 1999-04-15 Shaman Pharmaceuticals, Inc. Utilisation d'acide nordihydroguaiaretique pour la reduction du taux de triglycerides seriques et de la pression arterielle et pour le traitement du syndrome x
WO2000019989A3 (fr) * 1998-10-02 2000-11-09 Gruenenthal Gmbh Utilisation de derives de catechine comme inhibiteurs de proteinase
WO2003018036A1 (fr) * 2001-08-24 2003-03-06 Botanical Biotech Pty Ltd Composition permettant de detecter et/ou traiter des lesions et des tumeurs
EP1032387A4 (fr) * 1997-10-31 2004-02-11 Smithkline Beecham Corp Nouveaux complexes metalliques
WO2005070402A1 (fr) * 2004-01-08 2005-08-04 Newtree Industry Co., Ltd. Methodes et preparations de traitement de l'acne utilisant des composes de lignan
EP1845787A2 (fr) * 2005-01-27 2007-10-24 Erimos Pharmaceuticals LLC Formulations pour l'injection de butanes catecholiques, dont des composes ndga, a des animaux
WO2008042896A3 (fr) * 2006-10-02 2008-07-31 Erimos Pharmaceuticals Llc Dérivé de ndga tétra-substitué via des liaisons éther et des liaisons carbamate et leur synthèse et utilisation pharmaceutique
WO2009076449A1 (fr) * 2007-12-12 2009-06-18 North Carolina State University Procédés et compositions de traitement des virus du groupe pox
US7741357B1 (en) * 2006-04-17 2010-06-22 Johns Hopkins University Heterocyclic and carbonate derivatives of NDGA and their use as new anti-HIV and anti-cancer agents
SG169224A1 (en) * 2001-05-09 2011-03-30 Univ Johns Hopkins Method for treatment of tumors using nordihydrogualaretic acid derivatives
WO2014134202A1 (fr) * 2013-02-26 2014-09-04 Triact Therapeutics, Inc. Cancérothérapie
US9067875B2 (en) 2006-10-02 2015-06-30 Erimos Pharmaceuticals Llc Tetra-substituted NDGA derivatives via ether bonds and carbamate bonds and their synthesis and pharmaceutical use

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US4315916A (en) * 1978-07-18 1982-02-16 Lucille L. Filson Topical salve
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Cited By (30)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0288534A4 (fr) * 1986-10-28 1989-02-23 Chemex Pharmaceuticals Inc Compositions de butanes catecholiques avec du zinc.
EP0290442A4 (fr) * 1986-11-19 1990-07-03 Chemex Pharmaceuticals Inc Composes pharmacologiquement actifs et leurs melanges, compositions organiques et sels metalliques.
WO1990015603A1 (fr) * 1989-06-12 1990-12-27 Shiseido Company, Ltd. Composition antipruritique
AU628588B2 (en) * 1989-06-12 1992-09-17 Chuji Yanagawa Antipruritic composition
US5219847A (en) * 1989-06-12 1993-06-15 Shiseido Company, Ltd. Antipruritic composition
WO1995005156A1 (fr) * 1993-08-17 1995-02-23 Schering-Plough Healthcare Products, Inc. Compositions servant au traitement des cors, callosites et verrues
US5702694A (en) * 1993-08-17 1997-12-30 Schering-Plough Healthcare Products, Inc. Compositions for treating corns, calluses and warts
WO1999017761A1 (fr) * 1997-10-06 1999-04-15 Shaman Pharmaceuticals, Inc. Utilisation d'acide nordihydroguaiaretique pour la reduction du taux de triglycerides seriques et de la pression arterielle et pour le traitement du syndrome x
EP1032387A4 (fr) * 1997-10-31 2004-02-11 Smithkline Beecham Corp Nouveaux complexes metalliques
WO2000019989A3 (fr) * 1998-10-02 2000-11-09 Gruenenthal Gmbh Utilisation de derives de catechine comme inhibiteurs de proteinase
US6509368B1 (en) * 1998-10-02 2003-01-21 Gruenenthal Gmbh Use of catechol derivatives as proteinase inhibitors
AU766379B2 (en) * 1998-10-02 2003-10-16 Grunenthal Gmbh Use of catechol derivatives as proteinase inhibitors
SG169224A1 (en) * 2001-05-09 2011-03-30 Univ Johns Hopkins Method for treatment of tumors using nordihydrogualaretic acid derivatives
WO2003018036A1 (fr) * 2001-08-24 2003-03-06 Botanical Biotech Pty Ltd Composition permettant de detecter et/ou traiter des lesions et des tumeurs
CN101410100B (zh) * 2004-01-08 2015-05-06 黄在宽 利用木酚素类化合物治疗痤疮的方法及含有该化合物的组合物
CN101410100A (zh) * 2004-01-08 2009-04-15 黄在宽 利用木酚素类化合物治疗痤疮的方法及含有该化合物的组合物
WO2005070402A1 (fr) * 2004-01-08 2005-08-04 Newtree Industry Co., Ltd. Methodes et preparations de traitement de l'acne utilisant des composes de lignan
US8642646B2 (en) 2004-01-08 2014-02-04 New Tree Industry Co., Ltd. Method and composition for treating acne using lignan compounds
EP1845787A2 (fr) * 2005-01-27 2007-10-24 Erimos Pharmaceuticals LLC Formulations pour l'injection de butanes catecholiques, dont des composes ndga, a des animaux
EP1845787A4 (fr) * 2005-01-27 2011-02-16 Erimos Pharmaceuticals Llc Formulations pour l'injection de butanes catecholiques, dont des composes ndga, a des animaux
US8420692B1 (en) 2006-04-17 2013-04-16 The Johns Hopkins University Heterocyclic and carbonate derivatives of NDGA and their use as new anti-HIV and anti-cancer agents
US7741357B1 (en) * 2006-04-17 2010-06-22 Johns Hopkins University Heterocyclic and carbonate derivatives of NDGA and their use as new anti-HIV and anti-cancer agents
CN101547689B (zh) * 2006-10-02 2014-02-26 埃里莫斯医药品有限公司 通过醚键和氨基甲酸酯键四取代的ndga衍生物、它们的合成方法和药学用途
US8178527B2 (en) 2006-10-02 2012-05-15 Erimos Pharmaceuticals Llc Tetra-substituted NDGA derivatives via ether bonds and carbamate bonds and their synthesis and pharmaceutical use
JP2010505865A (ja) * 2006-10-02 2010-02-25 エリモス ファーマシューティカルズ エルエルシー エーテル結合およびカルバメート結合を介する四置換ｎｄｇａ誘導体ならびにそれらの合成および薬学的使用
WO2008042896A3 (fr) * 2006-10-02 2008-07-31 Erimos Pharmaceuticals Llc Dérivé de ndga tétra-substitué via des liaisons éther et des liaisons carbamate et leur synthèse et utilisation pharmaceutique
US9067875B2 (en) 2006-10-02 2015-06-30 Erimos Pharmaceuticals Llc Tetra-substituted NDGA derivatives via ether bonds and carbamate bonds and their synthesis and pharmaceutical use
WO2009076449A1 (fr) * 2007-12-12 2009-06-18 North Carolina State University Procédés et compositions de traitement des virus du groupe pox
WO2014134202A1 (fr) * 2013-02-26 2014-09-04 Triact Therapeutics, Inc. Cancérothérapie
US9834575B2 (en) 2013-02-26 2017-12-05 Triact Therapeutics, Inc. Cancer therapy

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