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WO1994018947A1 - Complexe protamine-adn de haute purete et son utilisation - Google Patents

Complexe protamine-adn de haute purete et son utilisation Download PDF

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Publication number
WO1994018947A1
WO1994018947A1 PCT/US1993/001542 US9301542W WO9418947A1 WO 1994018947 A1 WO1994018947 A1 WO 1994018947A1 US 9301542 W US9301542 W US 9301542W WO 9418947 A1 WO9418947 A1 WO 9418947A1
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WIPO (PCT)
Prior art keywords
aqueous
protamine
process according
dna complex
mixture
Prior art date
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PCT/US1993/001542
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English (en)
Inventor
Sharifa Karali
John K. Barberii
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Sharifa Karali
Barberii John K
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Filing date
Publication date
Application filed by Sharifa Karali, Barberii John K filed Critical Sharifa Karali
Publication of WO1994018947A1 publication Critical patent/WO1994018947A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans

Definitions

  • the present invention relates, generally, to a process for the preparation and use of chemotherapeutic agents.
  • the present invention relates to a process for the preparation of high purity
  • nucleoprotamine-DNA complex substances and a process for their use as an anti-tumor or anti-viral agent, including their use as an anti-AIDS agent. Additionally, research data exists to further suggest that certain aging
  • nucleoprotamine-DNA complex substances are also useful in a variety of other medical conditions, some of which are serious, in humans and other mammals. Elevated serum cholesterol levels have also responded favorably to treatment.
  • Nucleohistones have been generally known to the art to be closely associated with DNA, and research evidence exists to suggest that such substances protect DNA by wrapping about the double helix of the DNA in adult cells.
  • Hnilica, Lubomir S. The Structure and Biological Functions of
  • nucleoprotamines are physically associated with the DNA of embryonic tissue and are not found in adult cells.
  • protamines have overall positive charges and are in the basic range of pH. Additionally, both are bound to the negative charge of DNA, with known affinity constants, and both are shielded by the negative charge of DNA. Histones are known to be soluble in very dilute mineral acids, but insoluble in both very dilute mineral acids and mild aqueous NH 4 OH.
  • histones, and possibly protamines and protamine-like polypeptides exert a control function over DNA through a direct physical contact, or a lack thereof, at a myriad of sites along the DNA in the genome of all living cells.
  • This direct physical contact at the molecular level constitutes charge cloud interactions between the proteins and the deoxyribose background.
  • histones and protamines act as merely a protective wrapper for the cellular DNA and lack th expected variability in their amino acid sequence to control transcription of messenger RNA (mRNA).
  • DNA helices have a major and minor groove along the alpha-helix.
  • DNA bases appear to be in the bottom of the major groove, and the deoxyribose backbone in the bottom of the minor groove.
  • sequence specific proteins may attach at either the minor or major groove sites, along with histone and protamines or
  • protamine-like proteins to control transcription or DNA, has been widely accepted. See, Li, Hsueh Jei, Chromatin and Chromosome Subunits, Academic Press, New York (1977).
  • Simple systems for the control of DNA expression are also well known to the prior art, such as the Lactose Operon (LAC Operon) system of prokaryotic cells.
  • LAC Operon Lactose Operon
  • An analysis of this operon model illustrates the concept of repressors and inducers as being fundamental control systems for mRNA transcription. See, e.g., Kim, R., and S.H. Kim, "Direct measurement of DNA unwinding angle in specific interaction between lac operator and repressor.” Cold Spring Harbor Symp. Quant. Biol., 47: 481-484 (1983); Wang, J., M. D.
  • Galactose along with catabolite activator protein (CAP), cyclic AMP (c-AMP), and RNA polymerase are capable of acting as an inducer, displacing the LAC repressor protein from the operator site, presumably accessable through the major groove, binding with the promotor side, and allowing
  • glucose acts to block the formation of the active inducer complex and the cell's own heterogeneous repressor remains attached to the operator site, with no transcription of the LAC genes possible.
  • glucose controls the transcription of the LAC operon.
  • Repressors under this theory have a negative influence on transcription, and this is an important aspect of control which the invention focuses upon.
  • repressors may have developed, through evolution, as mutations, or acquired oncogenes, in very early unicellular promordial organisms.
  • a mutant, with an incomplete repressor, may have had a competitive, if not at least a metabolic advantage, if it could halt the
  • viruses may have developed their own cell-directed repressors, encoded into viral DNA or RNA, transcribed when the virus DNA infected the host DNA, or translated from viral RNA, and subsequently pre-packaged with the viral genetic material during lysogeny phase in prokaryotic cells.
  • C-rep The cell's repressor
  • the cell's repressor has evolved a very specific operator region to match its complementary operator site (e.g., only 27 base-pairs long, with some symmetry, in E. coli.), with matched base sequence by base pair to base pair in the operator region; a form of evolved primary structure, with a high rate constant of association (e.g., 7 x 10 9 m -1 sec -1 in E. coll.); and, other primary,
  • V-rep viral repressor
  • the general histological changes of tissue associates with the regression of a cell toward a cancer are known.
  • Such cells are less differentiated, tend to function and appear as embryonic tissues and have been described as chaotic in their metabolic pathways and metastatic without regard to their proper location.
  • control of protein synthesis means proper health for a cell. Conversely, the lack of control or proper regulation of protein synthesis results in aberrant
  • nucleoprotamine therapy states that the treatment of mammals with specifically timed collection of extracted nucleoprotamine and protamine-like proteins removes false repressors and false inducers, due to the lack of complete operon affinity in these heterogenetic proteins.
  • the allogenetic repressor is, in all likelihood, poorly physically bound to the operator region of the operon thereby physically preventing the attachment of the RNA polymerase to make the mRNA template of the protein.
  • protamine-DNA complexes After administered protamine-DNA complexes arrive in the repressed cell, there is a relative abundance of protamine, as compared with functional cellular DNA.
  • the actual substitution of protamine follows a simple competitive inhibition model where the success of replacing the foreign repressor protein is directly proportional to a high protamine-DNA/foreign repressor ratio.
  • the reaction is also influenced by the destruction of the allogenetic DNA of the original protamine-DNA complex, stopping the return of the protamine molecule to the allogenetic donor from the cell's heterogenetic DNA, thus making the reaction
  • an object of the present invention to provide an improved process for the production of high purity nucleoprotamine-DNA complex substances.
  • nucleoprotamine-DNA complex compounds as anti-tumor or anti-viral agents.
  • the foregoing and related objects are accomplished by a process in which high purity protamine-DNA complexes are prepared by collecting nucleoprotamines specific developmental stages of a life form, specifically, fertilized amphibian, egg by low temperature processing.
  • the process also includes the steps of sequential homogenization in a high concentration aqueous salt solution and a citric acid buffer, at a low pH of approximately 2.2, followed by ultracentrifugation to remove insoluble matter. These steps are then followed by an aqueous chloroform extraction to isolate protein and to remove lipids and lyophilization.
  • Single pass alumina chromatography is then used to separate each active protamine and protamine-like basic fraction. Dialysis against pure water removes excess salt, and
  • lyophilization increases concentration of each separated protamine and protamine-like protein.
  • Each isolate may the be reconstituted with 5% weight/volume heterologous or homologous DNA, in order to shield from charge toxicity.
  • Sterile filtration produces injection quality physiologic aqueous form.
  • a precipitation in sterile pure water, followed by lyophilization to remove water and to produce a solid form of the protamine-DNA complex obtained, is also recommended for dry preservation.
  • protamine-DNA complex Following isolation of the protamine-DNA complex, encapsulation of the prepared solid or aqueous protamine-DN complexes, in a specific carrier substance, may be
  • encapsulation carriers are known from prior art literature, such as, for example, liposomes and nanoparticles.
  • nucleo- protamine-DNA complex compound produced in accordance with the present invention as described hereinabove is
  • developmentally-timed nucleoprotamine has a utility for inhibiting tumor cell growth; inhibiting viral reproduction; and, regulating mammalian cellular metabolism by directly influencing DNA transcription at the macromolecular level - a phenomenon that may delay, or even reverse, the observed physiological changes we associate or attribute to aging.
  • the present invention concerns a process that allows for the maximum extraction of
  • nucleoprotamine from any fertilized egg source with the protamine being extracted at the proper time during
  • the eggs may be defrosted at room temperature and mixed with an equal volume of 4 M aqueous NaCl solution buffered to pH 2.2 with 1/10 volume 0.1 M sodium citrate.
  • Tissue homogenation was accomplished with a Brinkman Polytron homogenizer set on #6, as understood in the art, for 4 to 5 minutes until all the eggs are finely ground into a thick gray emulsion.
  • This thick emulsion is ultra- centrifuged at 15,000+ g's for 15 minutes in a refrigerated centrifuge.
  • the cloudy, gray supernate is then easily poured off the brown and black granular sediment.
  • This supernate contains the cytoplasm, without organelles, and nucleoplasm, with unbound nucleoprotamine released from its close relationship with DNA by the high concentration of salt and acidic pH.
  • Serum protein electrophoresis at this stage, further shows a crude, but relatively pure Beta electrophoretic range protein peak, i.e., the crude,
  • CPDNA protamine-DNA
  • Further processing includes chloroform extraction of protein. This requires the addition of 0.1 g of Na 2 CO 3 per 20 cc of supernate, stirred incubation at 50° C. for 30 minutes at an adjusted pH of 7 with glacial acetic acid, and the addition of an equal volume of chloroform with 0.1 volume amyl alcohol. The mixture is then shaken for 10 minutes and centrifuged to separation at 2,000 g. The topmost pure aqueous layer of the resultant three-layer liquid is discarded. The middle layer, being of chloroform- alcohol-protein is removed from the lower layer of
  • protamines identified by basic Isoelectric Focusing (IEF), with a pI of approximately 9.50. Further purification involves separating the purified protamines and protamine- like proteins into discrete fractions by alumina
  • the salt content in the foregoing procedure can be reduced to 0.09% (physiologic) saline by dialysis against pure water. Addition of DNA at approximately 5.0 mol percent causes microprecipitation during dialysis. This results in reconstitution of protamine-DNA via micro- precipitation of the free base protamine with the available 1:1 mole ration DNA and reduced toxicity.
  • the DNA used in reconstitution may be of heterogenetic or homogenetic origin, i.e., from the protamine donor tissue or the target tissue.
  • This reconstituted protamine-DNA complex (RPDNA) can then be encapsulated and directed more specifically to target tissues.
  • the crude protamine-DNA may be precipitated to form wispy white tendrils in sterile double distilled water. This precipitate is easily separated by repeated centrifugation at 15,000 g's and decanting off the
  • the wet precipitate can be crystalized in a lyophilizer at 0.001 torr and -40° C. until reduced to an amorphous light brown sticky material, with the consistency of coarse cotton candy.
  • protamine-DNA complex is readily absorbed across the
  • testing data is presented as tumor size, calculated volume and growth curves during in vivo testing against B16F10 murine melanoma in C57BL6 mice, modeled after the National Cancer Center
  • Protocols for limited cohort group testing.
  • V [(d 1 + d 2 )/4] 2 pI The volume of a given mouse's leg on Day 0 was
  • NTV net tumor volume
  • test group as opposed to the control group, generally had a smaller amount of net tumor volume.
  • the product produced in accordance with the present invention as described hereinabove can be used to treat humans afflicted with cancers.
  • the product is generally administered in the form of a mixture of aqueous fractions, or can be given as the precipitated protamine and DNA complex, in a powder type form.
  • the treatment of cancer was confirmed by a patient who had the condition known as adenocarcinoma of the prostate. After the treatment remission of the condition of
  • adenocarcinoma of the prostate was obtained, documented by reduction in Prostate Specific Antigen and clinical
  • the product disperses into a family of active polypeptides, small enough to be absorbed across a variety of cell membranes, including the gut, liver, and vascular channels.
  • the family of polypeptides is
  • the family of polypeptides gains access to the host cell DNA, and can strip away false repressors and inducers of tumor origin that are disrupting the normal expression of phenotype by the cancerous cell.
  • IP indicates intraperitoneal injection route
  • the methods of administering the product are disclosed as another feature of the invention.
  • the effective dose is in the range of 13 to 26 mg per kilogram body weight of recipient per day. In clinical trials, oral doses were given in this range once a day.
  • Administration may be by some suitable route including oral, rectal, nasal, topical (including buccal and
  • a typical once daily oral dose might consist of eight ounces of liquid carrier, such as water, with 1,000 to 2,000 mg of product dissolved in it.
  • the product might be administered in conjunction with other known medications such as Leuprolide, Buserelin, Goserelin, Narfarelin, Flutamide, Finasteride, Parazosin, Terazosin, Testolactone, Atamestane, or in conjunction with surgical modalities such as orchiectomy, or local radiation therapy.
  • other known medications such as Leuprolide, Buserelin, Goserelin, Narfarelin, Flutamide, Finasteride, Parazosin, Terazosin, Testolactone, Atamestane, or in conjunction with surgical modalities such as orchiectomy, or local radiation therapy.
  • the various formulations of the product may be prepared for use by any methods well known in the art of pharmacy.
  • Such formulations would generally include an acceptable carrier, in terms of being compatible with the polypeptide ingredients.
  • the formulations may not be limited to those suitable for oral, rectal, nasal, topical (including buccal and sublingual), vaginal or parenteral (including
  • the formulations may be presented in unit dose form for any route.
  • Oral formulations are not limited to pressed or molded tablet, capsules or caplets, aqueous solution, sustained release capsules, emulsified slurry, oral troche or lozenge, pastilles, mouthwashes, nanoparticles, or liposomes, all containing a predetermined amount of the polypeptide family constituents.
  • the proportions of each polypeptide may vary, depending upon the desired effect on the host tumor and experience of the administrator.
  • Formulations for topical administration may be made by mixing an appropriate carrier into ointments, creams, jels, or pastes.
  • Transderm patches may be made with current release technology.
  • Formulations for rectal administration may include suppository, and retention oil or liquid, and foam carriers.
  • Formulations for vaginal use include pessaries, tampons, creams, gels, pastes, foams, or sprays, all of the
  • Formulations for nasal administration include powdered precipitated polypeptides suitable for nasal inhalation, or gels, drops, sprays, packings, all of a compatible carrier.
  • Formulations for parenteral administration include sterile aqueous and non-aqueous injection solutions which may include antioxidants, buffers, bacteriostats, or
  • the formulations may be presented in unit dose or multidose containers, such as sealed vials, or ampules. Lyophilized preparations may require
  • formulations may include other agents or apparatus conventional in the art of the formulation in question, for example a nasal inhalator or transdermal patch dispersal system.
  • nucleoprotamine-DNA complex compound produced in accordance with the present invention as described hereinabove is used treating a human Acquired Immuno
  • Deficiency Syndrome When the new product was used for treatment of AIDS, it reduced the P-24 antigen and Beta 2 Microglobulins, all markets of HIV viral activity; reduced or slowed the loss of Human T lymphocytes from patients with Acquired Autoimmune Deficiency Syndrome (AIDS), and increased the T Helper (CD4) to T Suppressor (CD8) Ratio; reversed the weight loss of the wasting syndrome associated with AIDS; contributed to the sense of well-being of AIDS patients, by increasing energy levels, and reduced common complaints of fatigue, muscle aches, neuritis, and
  • the product disperses into a family of active polypeptides, small enough to be absorbed across a variety of cell membranes, including the gut, liver, and vascular channels.
  • the family of polypeptides is
  • the family of polypeptides gains access to the host cell DNA, and can strip away false repressors and inducers of HIV origin that are disrupting the normal expression of
  • the methods of administering the product are disclosed as another feature of the invention.
  • the effective dose is in the range of 1 to 15 m g per kilogram body weight of recipient per day. In clinical trials, oral doses were given in this range once a day, and mouse data indicated a half life of 24 to 48 hours.
  • Administration may be by some suitable route including oral, rectal, nasal, topical (including buccal and
  • a typical once daily oral dose might consist of eight ounces of liquid carrier, such as water, with 70 to 1,000 mg of product dissolved in it.
  • the product might be administered in conjunction with other known medications such as acyclovir, AZT, DDI, DDC, interferon, or other immunomodifiers or immunostimulants .
  • other known medications such as acyclovir, AZT, DDI, DDC, interferon, or other immunomodifiers or immunostimulants .
  • compositions of the product may be prepared for use by any methods well known in the art of pharmacy. Such formulations would generally include an acceptable carrier, in terms of being compatible with the polypeptide ingredients.
  • the formulations may not be limited to those suitable for oral, rectal, nasal, topical (including buccal and sublingual), vaginal or parenteral (including
  • the formulations may be presented in unit dose form for any route.
  • Oral formulations are not limited to pressed or molded tablet, capsules, or caplets, aqueous solution, sustained release capsules, emulsified slurry, oral troche or lozenge, pastilles, mouthwashes, nanoparticles, or liposomes, all containing a predetermined amount of the polypeptide family constituents.
  • the proportions of each polypeptide may vary, depending upon the desired effect of the host DNA, and experience of the administrator.
  • Formulations for topical administration may be made by mixing an appropriate carrier into ointments, creams, jels, or pastes.
  • Transderm patches may be made with current release technology.
  • Formulations for rectal administration may include suppository, and retention oil or liquid, and foam carriers.
  • Formulations for vaginal use include pessaries, tampons, creams, gels, pastes, foams, or sprays, all of the
  • Formulations for nasal administration include powdered precipitated polypeptides suitable for nasal inhalation, or gels, drops, sprays, packings, all of a compatible carrier.
  • Formulations for parenteral administration include sterile aqueous and non-aqueous injection solutions which may include antioxidants, buffers, bacteriostats, or
  • the formulations may be presented in unit dose or multidose containers, such as sealed vials, or ampules. Lyophilized preparations may require
  • formulations may include other agents or apparatus conventional in the art of the formulation in question, for example a nasal inhalator or transdermal patch dispersal system.
  • Patient 2 also again took the same oral regimen. He experienced a two percent reduction in the surface area of Kaposi's Sarcoma lesions. No side effects were documented.
  • P24 P24 Antigen, pg/ml

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Organic Chemistry (AREA)
  • Gastroenterology & Hepatology (AREA)
  • General Health & Medical Sciences (AREA)
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  • Engineering & Computer Science (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Genetics & Genomics (AREA)
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Abstract

Un complexe protamine-ADN de haute pureté est obtenu par un procédé en plusieurs étapes séquentielles, consistant essentiellement à recueillir et à traiter une nucléoprotamine obtenue à un stade embryonnaire d'une forme vivante par homogénéisation dans une solution aqueuse saline tamponnée pour obtenir un mélange, puis à enlever les matières insolubles du mélange obtenu après les étapes de collecte et de traitement précitées, à isoler la protéine et à éliminer les lipides du mélange par une extraction de la phase aqueuse par du chloroforme, à effectuer une dialyse de la protéine contre de l'eau stérile pour éliminer le sel en excès, à reconstituer la protéine à l'aide d'ADN hétérogène d'un tissu visé et à effectuer une filtration stérile pour obtenir un complexe aqueux protamine-ADN. Le procédé de traitement d'un patient présentant une tumeur consiste à administrer audit patient une dose appropriée du point de vue thérapeutique du produit de l'invention. Le procédé de traitement d'un patient humain souffrant du syndrome d'immunodéficience acquise consiste à administrer audit patient une dose appropriée du point de vue thérapeutique du produit de l'invention.
PCT/US1993/001542 1993-02-16 1993-02-16 Complexe protamine-adn de haute purete et son utilisation WO1994018947A1 (fr)

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US08/018,224 1993-02-16

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997004748A2 (fr) * 1995-08-01 1997-02-13 Advanced Therapies, Inc. Enveloppes virales artificielles renforcees pour l'apport de substances therapeutiques dans les cellules
WO2004098630A1 (fr) * 2003-05-12 2004-11-18 Nissan Chemical Industries, Ltd. Inhibiteur de troubles menopausiques
WO2005112885A2 (fr) 2004-05-12 2005-12-01 Baxter International Inc. Administration de microspheres d'oligonucleotides antisens pour induire une tolerance de cellules dendritiques pour le traitement du diabete de type 1 insulino-dependant
WO2005112894A1 (fr) * 2004-05-12 2005-12-01 Baxter International Inc. Production et administration de microsphères d’acide nucléique
US7374782B2 (en) 2000-10-27 2008-05-20 Baxter International Inc. Production of microspheres
CN113546056A (zh) * 2021-07-01 2021-10-26 温州医科大学附属第二医院（温州医科大学附属育英儿童医院） 一种针对阿霉素心脏及系统毒性解毒的仿生纳米保护剂制备方法及应用
CN113577039A (zh) * 2021-07-01 2021-11-02 温州医科大学附属第二医院（温州医科大学附属育英儿童医院） 鱼精蛋白压缩dna后被红细胞膜包裹构成的纳米颗粒的应用及其制备方法

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Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997004748A3 (fr) * 1995-08-01 1997-05-29 Advanced Therapies Inc Enveloppes virales artificielles renforcees pour l'apport de substances therapeutiques dans les cellules
WO1997004748A2 (fr) * 1995-08-01 1997-02-13 Advanced Therapies, Inc. Enveloppes virales artificielles renforcees pour l'apport de substances therapeutiques dans les cellules
US7374782B2 (en) 2000-10-27 2008-05-20 Baxter International Inc. Production of microspheres
WO2004098630A1 (fr) * 2003-05-12 2004-11-18 Nissan Chemical Industries, Ltd. Inhibiteur de troubles menopausiques
AU2005244842B2 (en) * 2004-05-12 2010-09-23 Baxter Healthcare S.A. Nucleic acid microspheres, production and delivery thereof
WO2005112885A3 (fr) * 2004-05-12 2006-02-09 Baxter Int Administration de microspheres d'oligonucleotides antisens pour induire une tolerance de cellules dendritiques pour le traitement du diabete de type 1 insulino-dependant
WO2005112894A1 (fr) * 2004-05-12 2005-12-01 Baxter International Inc. Production et administration de microsphères d’acide nucléique
EP2072040A1 (fr) * 2004-05-12 2009-06-24 Baxter International Inc. L'emploi thérapeutique des microsphères d'acide nucléique
WO2005112885A2 (fr) 2004-05-12 2005-12-01 Baxter International Inc. Administration de microspheres d'oligonucleotides antisens pour induire une tolerance de cellules dendritiques pour le traitement du diabete de type 1 insulino-dependant
EP2335689A1 (fr) * 2004-05-12 2011-06-22 Baxter International Inc. Méthode de production des microsphères d'acide nucléique
US9115357B2 (en) 2004-05-12 2015-08-25 Baxter International Inc. Delivery of AS-oligonucleotide microspheres to induce dendritic cell tolerance for the treatment of autoimmune type 1 diabetes
US9339465B2 (en) 2004-05-12 2016-05-17 Baxter International, Inc. Nucleic acid microspheres, production and delivery thereof
CN113546056A (zh) * 2021-07-01 2021-10-26 温州医科大学附属第二医院（温州医科大学附属育英儿童医院） 一种针对阿霉素心脏及系统毒性解毒的仿生纳米保护剂制备方法及应用
CN113577039A (zh) * 2021-07-01 2021-11-02 温州医科大学附属第二医院（温州医科大学附属育英儿童医院） 鱼精蛋白压缩dna后被红细胞膜包裹构成的纳米颗粒的应用及其制备方法
CN113546056B (zh) * 2021-07-01 2022-09-13 温州医科大学附属第二医院（温州医科大学附属育英儿童医院） 一种针对阿霉素心脏及系统毒性解毒的仿生纳米保护剂制备方法及应用
WO2023274105A1 (fr) * 2021-07-01 2023-01-05 温州医科大学附属第二医院（温州医科大学附属育英儿童医院） Procédé de préparation et application d'un agent de nanoprotection biomimétique pour lutter contre la cardiotoxicité et la toxicité systémique de la doxorubicine
US12102690B2 (en) 2021-07-01 2024-10-01 The Second Affiliated Hospital Of Wenzhou Medical University Preparation method and application of biomimetic nano-protectant for detoxifying dox-induced cardiac and systemic toxicity

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