SK172299A3 - DRUG DELIVERY DEVICES AND METHODS FOR TREATMENT OF VIRAL ANDì (54) MICROBIAL INFECTIONS AND WASTING SYNDROMES - Google Patents

Abstract

Novel drug delivery devices, methods and therapeutic compositions are described for treating viral and microbial infections and wasting syndromes in an animal, including a human patient. According to the invention, a polar compound such as dimethylformamide or dimethylsulfoxide is administered to a patient in need of treatment, preferably by a transdermal route. The invention further provides a vaccine prepared from antibodies harvested from the body of a patient treated by the method of the invention for a viral infection.

Description

Technical field

The present invention is directed to methods of treating animals suffering from viral or microbial infections, in particular retroviral infections such as HIV or weight loss syndrome, in particular weight loss syndrome associated with HIV infection or malignancy, by administering polar substances such as Ν, Ν'-dimethylformamide (DMF). ) or dimethylsulfoxide (DMSO). The invention further provides pharmaceutical compositions and compositions for the administration of a therapeutic agent consisting of polar substances, such as DMF or DMSO, for the treatment of animals affected by viral or other microbial infections or weight loss syndrome.

BACKGROUND OF THE INVENTION

Human immunodeficiency virus

Human immunodeficiency virus (HIV) induces persistent and progressive infection, which in most cases leads to the development of acquired immunodeficiency syndrome (AIDS) (Barre-Sinoussi et al., 1983, Science 220: 868-870, Gallo et al., 1984, Science 224: 500-503). There are at least two different types of HIV: HIV-1 (Barre-Sinoussi et al., 1983, Science 220: 868-870, Gallo et al., 1984, Science 224: 500-503) and HIV-2 (Clavel et al. , 1986, Science 223: 349-346, Guyader et al., 1987, Nature 326: 662-669). In humans, HIV replication is noticeably in populations of CD4 ⁺ T lymphocytes and HIV infection leads to depletion of this cell type and over time to immune deficiency, opportunistic infections, neurological dysfunction, neoplastic growth and eventually death.

HIV belongs to the retroviral family of lentiviruses (Teich et al., 1984, RNA Tumor Viruses, Weiss et al., Eds., CSHpress, pages 949-956). Retroviruses are small enveloped viruses that contain single stranded RNA in the genome and replicate via a DNA intermediate consisting of a virus encoded by reverse transcriptase, an RNA-dependent DNA polymerase (Varmus, H., 1988, Science 240: 1427-1439). Other retroviruses include, for example, oncogenic viruses, such as human T-cell leukemia virus (HTLV-I, -II, -III) and feline leukemia virus.

The HIV viral particle consists of a viral nucleus consisting in part of the p24 and p18 capsid proteins, together with the viral RNA genome and those enzymes that are essential for early replication processes. Myristylated forms an outer region around the viral nucleus that is surrounded by a membrane envelope derived from the infected membrane for alteration. Envelope surface glycoproteins synthesized as a single 160 kDa precursor protein that is cleaved by cellular proteases during the germination of the virus into two glycoproteins, gp41 and gp120. gp41 is a transmembrane glycoprotein and gp120 is an extracellular glycoprotein that remains non-covalently associated with gp41, possibly in a trimeric or multimeric form (Hammerskjold, M. and Rekosh, D., 1989, Biochem. Biophys. Acta 989: 269-280).

HIV, like other enveloped viruses, introduces genetic material into the host cell via the viral envelope and the target because the CD4 protein gag viral skin, the lipid cell HIV are membranes. HIV is a cell surface mediated fusion of viral targeting CD4 ⁺ cells, protein (CD4) acts as a cell receptor for HIV virus (Dalgleish et al., 1984, Nature 312: 767-768, Maddon et al., 1986, Cell 47: 333 -348). Viral entry into the cell is dependent on the binding of gp120 to the cellular CD4 receptor (Pal et al., 1993, Virology 194: 833-837; McDougal et al., 1986, Science 231: 382-385; Maddon et al., 1986, Cell). 47: 333-348), which explains the tropism of the virus to CD4 cells, while gp41 binds the envelope glycoprotein complex in the viral membrane. Binding of gp120 to CD4 induces conformational changes in viral glycoproteins, but this binding alone is not sufficient to lead to infection (reviewed in Sattentau and Moore, 1993, Philos. Trans. R. Soc. London (Biol.) 342 59-66).

Studies of HIV isolates have revealed heterogeneity in their ability to infect various human cell types (reviewed in Miedema et al., 1994, Immunol. Rev. 140: 35-72). Most extensively passaged laboratory strains of HIV clearly infect cultured T-cell lines and primary T lymphocytes, but not primary monocytes or macrophages. These strains are called T-tropical. HIV-infected individuals are more likely to occur in HIV-infected individuals during the late phases of AIDS (Weiss et al., 1996, Science 272: 1885-1886). Most primary HIV isolates (i.e., viruses that have not been extensively passaged in cultures) replicate effectively in primary lymphocytes, monocytes, and macrophages, but grow poorly on established T-cell lines. These isolates were designated as M-tropical. T- and M-tropism viral detreminants map to the third variable region of gp120 (V3 loop) (Choe et al., Cell 85: 1135-1148; Cheng-Mayer et al., 1991, J. Virol. 65: 6931-6941 Hwang et al., 1991, Science 253: 71-74; Kim et al., 1995, J. Virol., 69: 1755-1761; O'Brien et al., 1990, Nature 348: 69-73). Characterization of HIV isolates with different tropism, along with the observation that binding to CD4 cell surface protein alone is not sufficient to lead to infection, suggest that cell-specific cofactors might be required for CD4 to enter HIV into host cells. cells.

Treatment of HIV infection

HIV infection is pandemic and HIV-related diseases are a major global health problem. Despite considerable efforts to develop effective therapeutics, there are currently no anti-retroviral therapeutics against AIDS. In the development of such drugs, several stages of the HIV life cycle have been considered as possible targets for therapeutic intervention (Mitsuya H. et al., 1991, FASEB J. 5: 2369-2381). Many viral targets have been proposed for affecting the life cycle of HIV, although it is believed that affecting host cell proteins would have deleterious side effects. For example, virus-encoded reverse transcriptase was one of the focuses of drug development. Many drugs directed to reverse transcriptase have been developed, including 2 ', 3'-dideoxynucleoside analogs such as AZT, dd1, ddC and d4T, and have been shown to be active against HIV infection (Mitsuya, H. et al., 1991, Science 249: 15331544).

The new treatment regimen for HIV infection shows that combinations of anti-HIV drugs that target reverse transcriptase (RT), such as azidothymidine (AZT), lamivudine (3TC), dideoxyinosine (ddl), dideoxycitidine (ddC), used in combination with HIV protease inhibitors, have a greater effect (decrease by 2-3 logs) on the amount of virus compared to AZT alone (decrease by about 1 log). For example, recently, admirable results have been obtained with a combination of AZT, ddl, 3TC, and ritonavirus (Perelson, AS et al., 1996, Science 15: 1582-1586). However, long-term use of a combination of these chemicals is likely to lead to toxicity, particularly in bone marrow. Long-term cytotoxic therapy may also lead to suppression of CD8 ⁺ cells, which are essential for the control of HIV through killer cell activity (Blazevic, V. et al., 1995, AIDS Res. Hum. Retroviruses 11: 1335-1342) and through release of suppressive factors. , in particular the chemokinins Rantes, ΜΙΡ-Ια and MIP-Ιβ (Cocchi, F. et al., 1995, Science 270: 18111815). Another problem with long-term anti-retroviral therapy is the development of partial or complete resistance HIV mutations (Lange, JM, 1995, AIDS Res. Hum. Retroviruses 10: S77-82). It is believed that such mutations may be an inevitable consequence of antiviral therapy. The nature of the disappearance of wild-type viruses and the emergence of a mutated virus as a result of treatment, combined with a coincidence decrease in CD4 ⁺ cell counts, strongly suggest that the appearance of a viral mutant is a major underlying factor in AIDS therapy failure.

There are also attempts to develop drugs that can inhibit the entry of the virus into cells, the earliest stage of viral infection. Here, while the main focus has been on CD4, the cell surface receptor for HIV. For example, a recombinant soluble CD4 molecule has been shown to inhibit CD4 ⁺ T cell infection by some HIV-1 strains (Smith, DH et al., 1987, Science 238: 1704-1707). However, certain primary HIV isolates are relatively less susceptible to inhibition by the recombinant bio-CD4 molecule (Daar, E. et al., 1990, Proc. Natl. Acad. Sci. USA 87: 6574-6579). In addition, clinical trials with recombinant soluble CD4 have not given conclusive results (Schooley, R. et al., 1990, Ann. Int. Med. 112: 247-253; Kahn, JO et al., 1990, Ann. Int. Med. 112 Yarchoan, R. et al., 1989, Proc. Vth Int. Conf. On AIDS, p. 564, MCP 137).

Late stages of HIV replication, which include key viral specific processes of certain virus-encoded proteins, have also been suggested as possible targets for anti-HIV drugs. Late-stage processes are dependent on viral protease activity and drugs that inhibit this protease are being developed (Erickson, J., 1990, Science 249: 527-533).

Recently, CD8 + T-cell chemokinins have been included in the inhibition of HIV infection (Paul, W.E., 1994, Cell, 82: 177; Bolognesi, D.P., 1993, Semin. Immunol. 5: 203).

The chemokinins Rantes, ΜΙΡ-Ια and ΜΙΡ-1β, which are secreted by CD8 + T cells, have been shown to suppress the production of HIV p24 antigen in cells infected with HIV-1 or HIV-2 isolates in vitro (Cocchi, F. et al. 1995, Science 270: 1811-1815).

Therefore, these and other cytokinins could be useful in the treatment of HIV infection. However, the clinical outcome is questionable for all these and other candidate drugs.

Attention is also paid to the development of vaccines for the treatment of HIV infection. HIV envelope proteins (gp160, gp120, gp41) have been shown to be major antigens for anti-HIV antibodies present in AIDS patients (Barin et al., 1985, Science 228: 1094-1096). Therefore, to date, these proteins seem to be the most candidate candidates to act as antigens for the development of anti-HIV vaccines. Several groups have started using different portions of gp160, gp120 and / or gp41 as immunogenic targets for the host immune system. See, for example, Ivanoff, L. et al. U.S. Pat. 5,141, 867; Saith, G. et al., WO92 / 22,654; Shafferman, A., WO91 / 09,872; Formoso, C. et al., WO90 / 07,119. To date, HIV-targeted vaccines have been problematic in that viruses rapidly mutate, rendering many of these vaccines ineffective. However, the clinical results of these candidate vaccines are still far away.

Thus, although there is a great deal of effort to develop and test anti-retroviral drugs, effective non-toxic treatment is still needed.

Weight loss syndrome

Weight loss syndrome is a serious clinical problem characterized by a weight loss of more than 10% compared to baseline body weight and disproportionate body weight loss in relation to body fat (Weinroth et al., 1995, Infectious Agents and Disease 4: 76- 94; Kotler and Grunfeld, 1995, AIDS Clin. Rev. 96: 229275). Therefore, this weight loss is distinguished from starvation, where more body fat is lost than body cell weight (Kotler et al., 1985, Am. J. Clin. Nutr. 42: 1255-1265; Cahill, 1970, N. Engl. J. Med., 282: 668-675). Weight loss is associated with a variety of conditions including HIV infection, other infectious diseases, sepsis, cancer, chronic cardiovascular disease and diarrhea (Kotler et al., 1989, Am. J. Clin. Nutr. 50: 444-447;

Heymsfield et al., 1982, Am. J. Clin. Nutr.

Importantly, weight loss is a significant mortality in patients suffering from infections or cancer. In fact, weight loss is directly related to survival time in AIDS patients (Kotler et al., 1989, Am. J. Clin. Nutr. 50: 444-447).

The cause of weight loss syndrome in AIDS and other conditions is unclear and most likely multifactorial. Metabolic abnormalities, irregular hormone and cytokinin levels, and absorption disorders have been implicated in weight loss syndrome. Not all AIDS patients suffer from weight loss, which means that HIV alone is not the cause of weight loss. Most cases of HIV-related weight loss syndromes appear to be due to complications of AIDS, such as secondary infections and gastrointestinal diseases (Kotler and Grunfeld, 1995, AIDS Clin. Rev. 96: 229-275).

Current and potential therapies for weight loss syndrome include nutritional support, appetite enhancers

36: 680-690. food factors such as dronabinol megestrol acetate, anabolic therapies such as growth hormones and cytokinin inhibitors. However, mixed results were obtained with nutritional supplements and appetite enhancers in those patients who tended to accumulate only fat and not total body mass. The administration of growth hormones and cytokinin inhibitors is still being tested and may present a risk of side effects (Kotler and Grunfeld, 1995, AIDS Clin. Rev. 96: 229-275; Weinroth et al., 1995, Infectious Agents and Disease 4: 76-94) .

Therefore, weight loss treatment is critical to the survival and well-being of patients suffering from serious diseases such as cancer and AIDS. Therefore, there is a need for safe and effective therapy for weight loss syndrome associated with cancer, AIDS and other infectious diseases.

Properties of dimethylformamide and other polar substances

N, N'-dimethylformamide (DMF) (molecular formula C3H7ON) is a colorless, polar, hygroscopic liquid with low volatility and boiling point 152.5-153.5 ° C. It is freely miscible with water, alcohols and some hydrocarbons. DMF is generally used as a polar solvent and is well absorbed through the skin, by inhalation or after oral administration. DMF is rapidly metabolised, mainly in the liver and excretion occurs mostly in the urine. In rats, mice, hamsters and humans, the major metabolites of DMF are N-hydroxymethyl-N-methylformamide (HMMF), N-methylformamide (NMF) and N-acetyl-S- (N-methylcarbamoyl) cysteine (AMCC) as well as dihydroxymethylformamide ( DHMF) and N-hydroxymethylformamide (HMF). Unchanged DMF is excreted in the urine as a small fraction of the administered dose of DMF.

DMF has low acute dermal, oral and inhalation toxicity. It is considered to be a moderately irritating substance for the skin and eyes and easily passes through the skin. There is no indication that it has skin sensitizing properties. The major toxic effect of DMF and its metabolites is on the liver; DMF is known to cause reversible liver damage, which is associated with typical clinical complications, classical biochemical changes in blood, and the appearance of hepatocyte necrosis in liver biopsies. DMF is teratogenic but is not considered to be a mutagen or a carcinogen.

Viza et al. have reported that DMF and DMSO inhibit the in vitro replication of HIV and human herpes virus 6 (HHV6) in certain cultured cell lines (see Viza et al., 1990, AIDS Res. Hum. Retroviruses 6: 131-132; Viza et al., 1989, AIDS-FORSCHUNG 4: 349-352; Viza et al., 1992, Antviral res. 18: 27-38 and erratum at 19: 179).

DMF has been described as an in vitro differentiation agent for certain transformed cells in culture (see Koeffler, 1983, Blood 62: 709-721; Calabresi et al., 1979, Biochem. Pharmacol. 28: 1933-1941). When added to certain malignant cells in vitro, DMF has been reported to reduce their tumorigenicity following subsequent inoculation into nude mice (see Dexter, 1977, Cancer Res. 37: 3136-3140; Dexter et al., 1979, Cancer Res. 39 : 1020-1025). After intraperitoneal injection into nude mice, DMF and NMF have been shown to slow the growth of certain human tumor xenografts (see Van Dongen et al., 1989, Int. J. Cancer 43: 285-292;

Braakhuis et al., 1989, Head & Neck 11: 511-515; Van

Dongen et al., 1988, Acta Otolaryngol. 105: 488-493; Dexter et al., 1982, Cancer Res. 42: 5018-5022. However, the toxic side effects of formamide and its N-methyl derivatives in the mouse sarcoma allograft model led researchers to conclude that these substances are unlikely to prove therapeutically effective (see Čiarke et al., 1953, Proc. Sec. Exp. Biol Med 84: 203-207). Attempts to treat human tumor patients with DMSO led to the conclusion that an objective response was not achieved (see Spremulli and Dexter, 1984, J. Clin. Oncol. 2: 227-241). Oral administration of NMF to patients with head and neck cancer has been reported to result in hepatotoxicity without a beneficial response (see Vogel et al., 1987, Invest. New Drugs 5: 203-206) or with minimal activity (see Planting et al. , 1987, Cancer Treat Rep. 71: 1293-1294).

U. U.S. Patent No. 3,551,154 discloses the use of DMF as a penetration enhancer to promote transdermal absorption of topical drugs. U.S. Patent No. 4,855,294 discloses the use of glycerin to alleviate skin irritations induced by the use of DMSO and DMF as penetration enhancers to promote transdermal absorption of topical drugs. The use of DMSO as a penetration enhancer to support transdermal agents is discussed by J. Toxicol. Cut. & Ocular

Toxicol. 5: 167-177.

Citation or identification of any references in section 2 (or any other section) of this application shall not be construed as an admission that such reference is available as prior knowledge in relation to the present invention.

absorption in Woodford anti-viral and Barry, 1986,

SUMMARY OF THE INVENTION

The present invention is directed to methods, compounds and compositions for administering a therapeutic agent for treating an animal afflicted with a viral or other microbial infection, in particular a retroviral infection such as HIV. The invention further provides methods, compounds and compositions for administering a therapeutic agent for the treatment of an animal suffering from weight loss syndrome, such as weight loss associated with HIV infection or malignant disease.

According to the present invention, an animal in need of treatment is administered a compound consisting of N, N'-dimethylformamide (dimethylformamide, DMF), N-hydroxymethyl N-methylformamide (HMMF), N-hydroxymethylformamide (HMF), dihydroxymethylformamide (DHMF), N-acetyl S- ( N-methylcarbamoyl) cysteine (AMCC), N-methylformamide (NMF), dimethylsulfoxide (DMSO), formamide, acetamide, methylacetamide, dimethylacetamide, diethylacetamide, isopropylacetamide, diisopropylacetamide, N-acetylpiperidine, N, β (β); 1'-di (β-hydroxyethyl) acetamide, N-acetylmorpholine, acrylamide, propionamide, N-fluoromethyl-N-methylformamide, pyridine-N-oxide or any reagent selected from the group consisting of amides of the formula R ³ -CO-NR ¹ R ² , wherein R ¹ and R ² are independently selected from the group consisting of hydrogen, methyl, or unsaturated C 2 -C 3 hydroxylated alkyl halomethyl, saturated alkyl groups and groups; or

R ¹ and R ² are together selected from the group consisting of (CH ₂ ) ₄ , (CH ₂ ) ₅ 3 (CH ₂ ) ₂ O (CH ₂ ) ₂ ; and

R ³ is selected from the group consisting of hydrogen, methyl, and saturated or unsaturated C ₂ -C 3 alkyl groups. The therapeutic composition may consist of a mixture of two or more of the above.

The therapeutic regimen in an HIV-infected patient may advantageously combine a compound of the invention with one or more other agents effective in the treatment of HIV infection, including, but not limited to, selected from the group consisting of nucleoside analogue reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors and protease inhibitors in any desired combination.

The invention is also extended to vaccines prepared from antibodies that are obtained from an animal body after treatment of a viral infection with a compound of the invention.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides methods, compounds and compositions for administering a therapeutic agent for the treatment of viral or microbial infections. In an embodiment, the infection to be treated is an infection with retroviruses such as HIV, including an asymptomatic infection, a latent infection, and an infection accompanied by one or more AIDS-related symptoms and an infection accompanying clinical AIDS. Alternatively, the infection to be treated is any other viral or microbial infection, including rubella infection, herpes viruses such as human herpes virus 6, Epstein-Barr virus or cytomegalovirus, infection with any virus that has a capsid protecting the envelope, and any opportunistic infection associated with it. immune system such as opportunistic infection in HIV infected patients.

The present invention also provides methods, compounds and compositions for administering a therapeutic agent for treating or preventing any disease or disorder characterized by weight loss. Particular conditions that can be treated by the methods and compounds of the invention include, but are not limited to, weight loss associated with viral (e.g. HIV), bacterial or any other infection or sepsis, cachexia associated with malignant disease, chemotherapy, or radiation therapy, weight loss associated with chronic cardiovascular disease, weight loss caused by exposure of the patient to toxic or radioactive substances, and weight loss associated with diarrhea or other gastrointestinal diseases.

The subject to be treated may be any animal including, but not limited to, a monkey, a cow, a sheep, a cattle, a pig, a horse, a cat, a dog, a chicken, and others, preferably mammals, more preferably primates and most preferably an adult or a child, e.g. child weighing less than 3 kg. As used herein, the term patient means any animal in need of treatment with the methods or compounds of the present invention.

According to the present invention, the administered therapeutic agent consists of DMF, HMMF, HMF, DHMF, AMCC, NMF, DMSO, formamide, acetamide, methylacetamide, dimethylacetamide, diethyl acetamide, isopropylacetamide, diisopropylacetamide, N-acetylpiperidine, N- (β-hydroxyethyl) acetamide. , N'-di (β-hydroxyethyl) acetamide, N-acetylmorpholine, acrylamide, propionamide, N-fluoromethyl-N-methylformamide, pyridine-N-oxide, any reagent selected from the group consisting of amides of the formula R ³ -CONR ¹ R ² , wherein R ¹ and R ² are independently selected from the group consisting of hydrogen, methyl, halomethyl, saturated or unsaturated C 2 -C 3 alkyl groups and hydroxylated alkyl groups; and

R ³ is selected from the group consisting of hydrogen, methyl and saturated or unsaturated C ₂ -C ₃ alkyl groups, or any agent selected from the group consisting of amides of the formula R ³ -CO-NR ¹ R ² , wherein:

R ¹ and R ² are independently selected from the group consisting of hydrogen, methyl, halomethyl, saturated or unsaturated C 2 -C 3 alkyl groups and hydroxylated alkyl groups; or

R ¹ and R ² are together selected from the group consisting of (CH ₂ ) ₄ , (CH ₂ ) ₅ and (CH ₂ ) ₂ O (CH ₂ ) ₂ ; and intradermal, intraperitoneal,

R ^J is selected from the group consisting of hydrogen, methyl, and saturated and unsaturated C2-C3 alkyl groups.

In one particular embodiment, at least one of R ¹ and R ^{2 is} methyl. In another particular embodiment, the at least one of R ¹ and R ² C _x -C 3 fluorinated alkyl group. The therapeutic compound may consist of a mixture of a mixture of two or more of the above. Particularly preferred is a compound formed by DMF.

In addition to the compounds of the invention, the therapeutic regimen of an HIV-infected animal may advantageously comprise one or more agents effective in the treatment of HIV infection, e.g. one or more nucleoside analogue reverse transcriptase inhibitors such as zidovudine (AZT, ZDV), zalcitabine (ddC), didanosine (ddl), lamivudine (3TC), stavudine (d4T), one or more non-nucleoside reverse transcriptase inhibitors, nevirapine, delavirdine, loviride, atevirdine, pyridinone, one or more protease inhibitors such as saquinavir, indinavir, ritonavir, nelfinavir, or any combination of the above or other anti-HIV therapeutic agents. The compound of the present invention and the other anti-HIV therapeutic agent (s) may be administered simultaneously, sequentially or in treatment cycles according to the desired treatment protocol.

The compound of the invention may be administered by any desired enteral or parenteral route, including, but not limited to, transdermal, subcutaneous, intramuscular, intravenous, intranasal, epidural, intralymphatic or oral route. The agents may be administered by any conventional means, by infusion or injection, absorption through an epithelial mucocutaneous backing layer (e.g., oral, e.g.

or gastric, intestinal or rectal mucosa) and may be administered with other biologically active agents.

Administration may be systemic or local. In addition, it can

It may be desirable to deliver the pharmaceutical compound of the invention to the central nervous system by any suitable means, including intraventricular and intrathecal injection; administration by intraventricular injection may be facilitated by an intraventricular catheter attached to the reservoir, e.g. to the Ommaya stack. Pulmonary administration, e.g. using an inhaler or nebulizer containing an aerosol substance. If appropriate, two or more routes of administration may be used simultaneously, sequentially or in treatment cycles according to any therapeutic protocol.

In a particular embodiment, it may be advantageous to administer the compound of the invention to an area in need of treatment; this can be achieved, for example, but not limited to, by injection, via a catheter, a suppository, or an implant, said implant being a porous, non-porous or gelatinous material, including membranes such as sialolytic membranes and fibers.

In another embodiment, a compound of the invention may be administered in a vesicle, particularly a liposome (see Langer, Science, 249: 1527-1533 (1990); Treat et al., Liposomes in the Therapeutic of Infectious Disease and Cancer, LopezBerestein and Fidler (eds). Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., Pp. 317-327, see generally ibid.).

In yet another embodiment, the compound of the invention may be administered by a controlled release system.

In one embodiment, a pump may be used (see Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng. 14: 201 (1987); Buchwald et al., Surgery 88: 507 (1980); Saudek et al., N. Eng J Med 321: 547 (1989)). In another embodiment, polymeric materials can be used (see Medical

Applications of Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton, Florida (1974); Controlled Drug Bioavailability, Drug Product Design and Performance, Smolen and Bali (eds.), Wiley, New York (1984); Ranger and Peppas, J. Macromol. Sci. Rev. Macromol. Chem. 23: 61 (1983); see also Levy et al., Science 228: 190 (1985); During et al., Ann. Neurol., 25: 351 (1989); Howard et al., J. Neurosurg. 71: 105 (1989)]. In yet another embodiment, controlled release systems may be located near the therapeutic target, requiring only a portion of the systemic dose (see, e.g., Goodson, Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138 (1984)).

Other controlled release systems are discussed in Langr's review article (Science 249: 15271533 (1990)).

The present invention also provides pharmaceutical compositions. These compositions comprise a therapeutically effective amount of a compound of the invention and a pharmaceutically acceptable carrier. In a particular embodiment, therapeutically acceptable means a term that has been approved by a federal or state or other government regulatory agency as set forth in U.S. Pat. pharmacopoeia or other generally recognized pharmacopoeia for animals, more specifically humans. The term carrier refers to a solvent, adjuvant, excipient or vehicle with which a compound of the invention is administered. The pharmaceutical carriers can be sterile liquids, e.g. water and oils, including petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Water is the preferred carrier when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be used as liquid carriers, especially in injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, skimmed milk powder, glycerol, propylene glycol, water, ethanol and the like. If preferred, the pharmaceutical composition may also contain minor amounts of wetting, emulsifying or pH buffering agents. These compositions may take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained release formulations and the like. The composition may be formulated as a suppository with traditional binders and carriers such as triglycerides. Oral compositions may contain standard carriers such as pharmaceutical forms of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, and the like. Examples of suitable pharmaceutical carriers are described in Reminton's Pharmeceutical Sciences by E.W. Martina. Such compositions will contain a therapeutically effective amount of the therapeutic agent, preferably in purified form, together with a suitable amount of carrier to provide the form for proper administration to the patient. The composition should suit the mode of administration.

The compounds of the present invention may be administered transdermally. In one embodiment, the compound of the invention is applied directly to the skin. In another embodiment, the compound of the invention is applied to a reservoir (e.g., a cotton pad, a synthetic polymer such as Teflon ™ or any suitable adsorbent) that is applied to the skin, preferably under a closed bandage. In a particular embodiment, the compound of the invention is applied to the skin by means of dermal patches. The concentration of the active therapeutic agent in the composition that is applied to the skin, contained or adsorbed to the reservoir, or contained in the dermal patch may be 10% to 100%, preferably at least% and more preferably at least 90%.

In a preferred embodiment, the polar substance, e.g. DMF, administered transdermally using any suitable means of administering the therapeutic agent, e.g. by applying one or more dermal patches. The dermal patch may optimally consist of an outer support portion, a reservoir such as an adsorbent impregnated with a polar compound of the invention and a permeable membrane in contact with the skin. The support portion can be any material, such as a natural or synthetic polymer, that resists the chemical action of a polar substance. High density polyethylene backing is particularly preferred. The adsorbent may be a colloidal substance, e.g. diatomaceous earth or colloidal silica. The permeable membrane may be of any material that chemically resists the polar material and may optionally be supplied with pores. In a preferred embodiment, the Teflon ™ permeable membrane has a pore diameter of about 0.1 µm, a diameter of about 0.5 µm, or a diameter in the range of about 0.1 µm to 0.5 µm. The patch may be adhesive alone or may be in contact with the skin by means of an applicator such as a wrap or bandage, including, without limitation, an elastic or adhesive bandage; Elatoplast ™ is suitable for this purpose. Preferably, the patch contains a greater amount of a polar substance than is to be administered to the skin of the treated patient; e.g. the patch may contain about 50% more polar compound than is to be administered. The patch may be of various sizes and shapes and may, for example, be in the form of a disc having a diameter of approximately 9 cm.

In one embodiment, the patch comprises a polar agent such as DMF and at least one other pharmacologically active agent, e.g. anti-irritant glycerin. In another embodiment, the patch is a polar substance such as DMF and is not a skin lubricant and a moisturizer such as KY ™ jelly. The patch is then removed from the backing material and applied so that the permeable membrane is in contact with the prepared skin surface. The patch may be held in place by the applicator. After the desired dosing period, the patch is removed. After removal of the patch, the treated area may be thoroughly washed with a mild soap to remove residual polar substance.

The compounds of the invention may be administered at any desired interval, for example, once or twice every three weeks; once, twice or three times a week; every other day or daily. Preferably, the compound of the invention is administered at a dose that results in a plasma concentration of about 2 to 200 mg / L, more preferably 100 to 200 mg / L, even more preferably 150 mg / L of the active ingredient of the formulation such as DMF. A level of 100 to 150 mg / l or 150 to 200 mg / l DMF is particularly preferred. As used herein, the term ppm refers to a part of a million by weight and in practice is equivalent to mg / L.

For transdermal administration of a polar substance, the absorption level is determined by the skin of the test subject. After human skin exposure, DMF is absorbed at a steady rate of approximately 9.4 mg / cm ² per hour. (see Mraz and Nohova, Occup. Env. Health 64: 85-92). Likewise, the desired level of absorption can be achieved by controlling the area of the skin area exposed to the therapeutic agent as well as determining the area of each patch and the number of patches applied to the skin. For example, two patches with a diameter of 9 cm will expose the polar substance to a total skin area of 127 cm ² ; for DMF, this will result in an absorption rate of about 1.2 g DMF per hour. A starting dose of 15 mg / kg DMF is particularly preferred.

In one embodiment, a 72 kg patient is initially treated with DMF by administering two dermal patches present with no additional pharmacologically active agent. In another embodiment, the patch comprises a polar substance such as DMF and there is no other pharmacologically active substance present that is capable of being systemically absorbed through the skin or no other pharmacologically active substance is present in an amount that is systemically effective after transdermal absorption. In yet another embodiment, the patch comprises a polar substance such as DMF and no other antiviral agent is present, e.g. acyclovir or arildon.

Thus, the present invention provides a dermal patch comprising a polar substance such as DMF in an amount effective to treat a viral or microbial infection (e.g., HIV infection) in human adult or pediatric patients. The present invention further provides a dermal patch comprising a polar substance, such as DMF, in an amount that is effective in treating weight loss syndrome in human adult or pediatric patients. In one embodiment, the patch comprises at least 0.25 g of a polar material such as DMF, preferably at least 0.5 g, more preferably at least 1.0 g and even more preferably at least 5.0 g, e.g. 5.0 to 15.0 g of a polar material such as DMF.

In order to minimize the loss of polar material by evaporation, the patch may preferably be stored in a sealed package as a sealed polymer bag. If desired, each patch can be individually sealed for convenient use. Preferably, the patch may be cooled before use, e.g. to 4 ° C to reduce the loss of polar material by evaporation. Preferably, the patches are prepared for 24 hours prior to use and are stored in a sealed package at 4 ° C; however, the patches are stable for one or more weeks when stored at 4 ° C in a sealed package. Before applying the patch, it is preferable to wash the skin with a mild soap and water, rinse to remove the soap residue, dry thoroughly and then hydrate with a suitable

Treatment of HIV infection by transdermal administration of N, N'-dimethylformamide (DMF)

HIV-1 infected patients were treated with dimethylformamide (DMF) by applying skin patches impregnated with dimethylformamide gel to the patient's body. N-acetylcysteine-glutathione and / or necessary phospholipids were administered to the patient (orally or intravenously) daily at a dose of 250-300 mg as a liver function enhancer. Glutamine may be administered instead or in conjunction with it as a liver function enhancer.

Two skin patches have been applied to different parts of the body, e.g. forearm. Each skin patch contained 7.06 g of DMF gel (92.5% w / w) and colloidal silica (7.5% w / w). The gel served to prevent the leakage of liquid DMF through the patch. The patches were made for a maximum of 12 hours before use as DMF evaporates rapidly. The intended level of DMF in the patient's blood was 100 ppm. For a patient weighing about 60 kg, an amount of about 14 g is required over a 12 hour period to reach a level of 100 ppm. Based on studies by Mráz and Nohová, the area to absorb this amount is about 127.2 cm ² . To achieve a blood level of 100 ppm, about 1.272 g of DMF must be absorbed per hour, so each patch must contain about 7.064 g of DMF to deliver the required amount at a surface area of 6.36 cm ² (each patch). The absorption rate of DMF is 9.4 mg / cm ² / hour. This treatment theoretically delivers 125-135 ppm, but 100 ppm has been achieved due to DMF evaporation. Absorption capacity varies from patient to patient depending on factors such as skin type and skin thickness. To achieve the desired DMF level in the patient, plasma concentrations were monitored in each patient and the treatment was adjusted according to the DMF levels in each patient. Fig. 1 shows a diameter of 9 cm over one hour, resulting in an initial absorption dose of about 1.2 g DMF, corresponding to a dose of 16.7 mg / kg. The number of patches may be increased or decreased, and a longer or shorter initial exposure interval may be used, depending on the body weight of the subject being treated. This initial dose may be repeated at any desired interval as described above, and is preferably administered at weekly intervals. Preferably, the patient is monitored for toxicity at least 72 hours after DMF administration, for example by monitoring serum or plasma levels of enzymes such as aspartate aminotransferase (AST), alanine aminotransferase (ALT), γ-glutamyl transferase (yGT) and alkaline phosphatase, proteins such as albumin. , or substances such as conjugated or unconjugated bilirubin. Preferably, serum or plasma levels of AST and ALT do not exceed five times the upper limit of normal as determined by the reference range of the laboratory and the respective population; more preferably, serum or plasma levels of AST and ALT do not exceed three times the upper limit of normal, and most preferably serum or plasma levels of AST and ALT are not elevated above normal or above levels observed prior to treatment. The dose of DMF may be increased by applying the patches for a longer period of time; in one embodiment, the patches are applied for two hours, then for four hours, then for six hours, etc .; in another embodiment, the sequential exposure time is doubled. Preferably, the patient is monitored prior to each dose increase for any signs of toxicity. If desired, the dose may be increased at weekly intervals. In this way, the dose may be increased up to a calculated dose of about 150 mg / kg; or until the desired upper plasma is reached: a level of, for example, 150-200 mg / L; or until the exposure time is six to eight hours. Dose increases above 240 mg / kg should be used with caution.

plasma DMF levels in four patients treated with 2 dermal DMF patches for 8 hours. Fig. 2 shows plasma DMF levels in three patients treated with 2 dermal DMF patches for 6 hours.

Thus, the patches were impregnated with about 7.064 g of a gel with DMF and silica. Each patch was applied for 12 hours, either once a week for 12 weeks or twice a week for 6 weeks.

Blood tests showed an increase in the number of CD4 T cells from 350 to 1000 in certain patients and a rapid reduction in PCR (polymerase chain reaction) (viral load) from 120000 to 500 / ml within 3 weeks after only three treatment cycles. Fig. 3 shows a serial quantitative PCR measurement of the HIV-1 viral load in a patient before treatment and after three cycles of treatment with 2 dermal patches with DMF for 12 hours. The PCR tests performed were the Roche Amplicor HIV monitor. A viral load of less than 500 / ml plasma is considered undetectable.

Some treated patients had severe acne or showed symptoms of rubella before treatment. When treated with DMFF so that the DMF levels in the patient's blood were 50 to 100 ppm, the symptoms of rubella and severe acne disappeared within 7 days.

Before treatment with dimethylformamide, the patient underwent an overall baseline clinical and psychological evaluation. The evaluation provided basal levels of the patient's biochemical and hematological data. Detailed virological serum tests (HIV-1) were also performed to determine the total amount of virus in the patient and were performed weekly or by treatment.

During the treatment period, the concentration of DMF in the patient's blood was determined every hour. Each morning he was given an intravenous cannula to collect blood samples and was left open with an infusion of normal saline at a rate of 20 ml per hour. Monitoring of the active metabolite AMCC (e.g., by urine sampling after 4 hours), derived from DMF, was performed daily. Subsequent DMF applications were adjusted in accordance with the measured changes in blood DMF levels that resulted from changes in absorption variables, and complete hematological and biochemical profiles were performed daily to detect any changes in liver function. Full clinical and psychological evaluations were also performed daily.

Virological serum calculations were also performed daily to determine the total number of viruses in the body and to monitor the improvement of the patient's immune status (CD4 helper T-cells) and prognostic factors. Serological calculations were performed based on p24 antigen and quantitative PCR or other preferred methods. CD4 cell counts and β-2 macroglobulin were also evaluated weekly to specifically monitor the patient's immune status and prognosis. All clinical and laboratory data were delivered to a central data system to facilitate a rapid response to any harmful changes in order to limit treatment so that the maximum clinical effect is achieved and possible side effects are minimized.

Tests included:

a) serum: Na, K, Cl, CO2, urea, urate, creatinine, Ca,

Mg, phosphate, total and conjugated bilirubin

b) hematology: hemoglobin, red blood cell count, hematocrit, MCV, MCH, MCHC, RDW

c) serum protein electrophoresis: total proteins, albumin, total globulin, alpha-globulin, alpha2globulin, betaglobulin, gamaglobulin

(d) white blood cell analysis: white blood cell count, absolute neutrophil count, lymphocyte count, monocyte count, eosinophils and basophils

(e) liver enzymes: alkaline phosphatase, gamma GT;

ALT (SGPT); AST (SGOT); LDH

f) other: cellular traits, PCR, beta2-macroglobulin, p24 antigen, C-reactive protein, CK-MB concentration

g) blood analysis for DMFF level

h) urine analysis for AMCC levels

DMF appears to act as at least one of the reverse transcriptase inhibitors and protease inhibitors. In vitro assays have been performed and the dissolution properties of DMF appear to break down viral particles, e.g. capsid.

BRIEF DESCRIPTION OF THE DRAWINGS

The present invention may be elucidated by reference to a detailed description of the invention, exemplary embodiments of the invention, and the accompanying drawings, in which:

Figure 1 shows plasma DMF concentration as a function of time in 4 patients treated with 2 dermal patches for 8 hours, Figure 2 shows plasma DMF concentration as a function of time in 3 patients treated with 2 dermal patches for 6 hours, Figure 3 shows viral load by quantitative PCR in patients treated

DMF; two DMF patches were placed on the forearm skin for 12 hours on days 0.8 and 13 (indicated by arrows), Figure 4 shows the general condition of HIV-infected patients before and after treatment with transdermal DMF as assessed according to the Karnovsky scale of performance, see text, section 7, about details.

HIV-1, measured by transdermal

DETAILED DESCRIPTION OF THE INVENTION

Example

Treatment of HIV infection by transdermal administration of N, N'-dimethylformamide (DMF)

A preliminary study was conducted to determine the efficacy of transdermal DMF in the treatment of HIV infected patients. Informed consent was obtained from each patient. The seropositive condition was examined by Western blot using a commercial kit for the detection of anti-p24 antibodies (Abbott Diagnostics) and the presence of HIV-1 was documented by quantitative PCR using a commercial kit (Roche Amplicor).

DMF obtained commercially (Sigma-Aldrich) and in plasma samples was analyzed by mass spectroscopy / gas chromatography (GC / MS) using a Varian 9600 gas chromatograph, an OV351 column, a capillary carbos wax-PEG column and an ion trapping Finnegan Mat ITS40 detector. The operating parameters were as follows: GC temperature program: 60 ° C for 1 minute followed by a 9.4 ° C / min temperature gradient. for 20 minutes, MS ionization method: electron impact, mass range: 40-80 mass units, 1 image / sec, peak threshold: 3 calculations / sec, background mass: 69 mass units. A dimethylacetamide stock solution was used as internal standard. All samples were extracted with an organic solvent containing an internal standard, allowed to precipitate for 30 minutes in a refrigerator, and centrifuged at 3000g for 5 minutes before being transferred to GC ampoules for injection. The retention time of the DMF peak was 3.26 minutes relative to the internal standard; the calibration curve showed a correlation of 0.98. DMF quantitation was linear up to 100 mg / ml and the lower limit of detection was set at 0.5 mg / ml based on a 3: 1 signal to background ratio.

Dermal patches with a diameter of 9 cm were used within 12 hours of manufacture. Each patch had a support portion of high density polyethylene (0.245 g), a permeable membrane of Teflon ™ (pore size 0.2 µπι; 0.268 g) for placement against the skin and containing 0.573 g of colloidal silica impregnated with 7.067 g DMF between the support and a permeable membrane. The patches were visually inspected to confirm the absence of penetration and weighed on an analytical balance to confirm a deviation of less than 10% from a total weight of 8.153 g. The patches were applied to the forearm skin and secured with an Elatoplast ™ bandage. Either one or two patches were used at the initial dose as determined from an estimated skin patency rate of 9.4 mg / cm ² / hr. and the patient's body weight.

A phased dose of DMF was sampled every two hours to determine peak plasma levels of DMF. Patients were clinically monitored for any toxic side effects and were kept under overall biochemical control while the dose was increased to achieve a maximum plasma level of 100-120 ppm. Once the correct dose was established, transdermal DMF was used once a week. The initial evaluation included a daily determination of:

(a) vital signs of body weight

(b) the clinical list and the Carpathian score

c) total blood cell count and erythrocyte sedimentation rate

d) serum urea, creatinine, glucose, sodium, potassium, ALT, AST, alkaline phosphatase and total bilirubin

e) CD4 ⁺ and CD8 ⁺ cell count and CD4 ⁺ / CD8 ⁺ ratio using a Coulter instrument

f) quantitative estimates of HIV-1 burden using PCR (Roche Amplicor) and analysis of anti-p24 antibodies (Abbott)

g) urine analysis (Dipstix)

At each weekly visit, the patient was evaluated for adverse reactions, nearly all of these tests were repeated and blood and urine samples were taken to determine DMF and its metabolites.

Patient 1 (ADF) started the protocol in relatively good health, basically complaining of hand and foot pain and inability to sleep. He was given two DMF patches once a week, with an average exposure time of 8 hours. The average weekly dose of DMF was 6.11 g and the resulting mean maximum plasma DMF level of 75 mg / L. After 9 weeks, the number of CD4 ⁺ T cells increased from 140 to 640 cells / μΙ and the viral load measured by PCR was reduced from 250,000 to 50,000 copies / ml. After 10 weeks, the patient's weight increased from 81.9 to 96.0 kg, the patient felt clinically comfortable and no longer complained of limb pain.

Patient 2 (AM) entered a protocol complaining of loss of strength, inability to sleep, hand and foot pain, and herpetic ulcers in his mouth. He received one DMF patch per week with an average exposure time of 8 hours. The average weekly dose of DMF was 7.12 g and the resulting mean maximum plasma DMF level of 125 mg / L. After 9 weeks, the number of CD4 ⁺ T cells increased from 460 to 720 cells / μΙ and the viral load measured by PCR was reduced from 29,000 to 13,000 copies / ml and the patient's weight increased from 58.4 to 63.0 kg. Herpetic ulcers were cured, limb pain disappeared and the patient appeared clinically good.

Patient 3 (SM) started a protocol with overt clinical AIDS and complained of respiratory problems. He was given two DMF patches once a week with an average exposure time of 8 hours. The mean weekly dose of DMF was 8.17 g and the resulting mean maximum plasma DMF level of 121 mg / L. After 7 weeks, the number of CD4 ⁺ T cells increased from 39 to 138 cells / μΙ and the viral load measured by PCR was reduced from 222,000 to 16,000 copies / ml and the patient's weight increased from 74.2 to 100.0 kg. The patient's appetite improved, respiratory problems were resolved and the patient appeared clinically good and began to exercise again.

Patient 4 (EM) initiated a protocol with secondary infection (including herpes), anemia, diarrhea and acne. He was given two DMF patches once a week with an average exposure time of 8 hours. The mean weekly dose of DMF was 7.33 g and the resulting mean maximum plasma DMF level of 90 mg / L. After 18 weeks, the number of CD4 ⁺ T cells increased from 24 9 to 450 cells / μΙ and the viral load measured by PCR was reduced from 13,000 to 400 copies / ml and the patient's weight increased from 81.5 to 90.4 kg. The patient appeared clinically well and had no active health problems.

Patient 5 (SV) began the protocol by complaining of anorexia, forgetfulness, abdominal pain, and severe fatigue that prompted him to consider selling his share of the business. He was given two DMF patches once a week with an average exposure time of 6 hours. The mean weekly dose of DMF was 3.75 g and the resulting mean maximum plasma DMF level of 67 mg / L. After 5 weeks, the number of CD4 ⁺ T cells increased from 354 to 396 cells / μΙ and the viral load measured by PCR was reduced from 156,000 to 13,000 copies / ml and the patient's weight increased from 56.0 to 58.0 kg. The patient was clinically doing well, got his partner's actions and started running the business himself.

Patient 6 (VV) began the protocol by complaining of secondary infections (including herpes), ataxia, and numbness of the left hand and left side of the face. He was given two DMF patches once a week with an average exposure time of 8 hours. The mean weekly dose of DMF was 8.25 g and the resulting mean maximum plasma DMF level of 110 mg / L. After 19 weeks, the number of CD4 ⁺ T cells increased from 260 to 450 cells / μΙ and the viral load measured by PCR was reduced from 120,000 to 24,000 copies / ml and the patient's weight increased from 75.4 to 84.6 kg. Secondary infections disappeared and the patient appeared clinically good.

Patient 7 (AJF) started the protocol seriously ill. Patients initial CD4 ⁺ T-cell status was 29 cells / μΙ and the initial PCR measured viral load was 1.156,000 copies / ml. Treatment was with one patch with an average exposure time of 4 hours. The mean weekly dose of DMF was 4.60 g and the resulting mean maximum plasma DMF level of 100 mg / L. After the first treatment, the number of patient CD4 ⁺ T cells decreased to 14 cells / μΙ. Treatment was administered daily for the first five days and then continued at weekly intervals. After 9 treatment cycles, the number of patient CD4 ⁺ T cells increased to 39 cells / μΙ and the viral load measured by PCR was reduced to 9,000 copies / ml and the patient's weight increased from 46.5 kg (at the start of DMF therapy) to 49 0 kg. the patient felt comfortable and returned to full-time employment.

Patient 8 (MS) initiated a protocol with severe herpetic lesions on the lower back and genitals. He was given two DMF patches once a week with an average exposure time of 8 hours. The average weekly dose of DMF was 6.24 g and the resulting mean maximum plasma DMF level of 130 mg / L. After 8 weeks, the CD4 ⁺ T-cell count increased from 200 to 240 cells / μΐ and the viral load measured by PCR was reduced from 1,200,000 to 250,000 copies / ml and the patient's weight increased from 48.1 to

52.2 kg. Herpetic lesions have completely disappeared.

Two patients were excluded from the tests, one because of alcohol dependence, the other because of viral hepatitis

B.

Most patients have experienced mild local irritation in the application area after patch removal; the skin at the application site had a maculopapular appearance, probably due to intense hydration under the patch. In one case, mild pimples were formed which disappeared within 24 hours and did not cause significant problems to the patient. Most patients experienced mild transient nausea, usually on the third day of treatment, which gradually decreased during the treatment protocol; one patient reported moderate transient nausea. Four patients exhibited a transient increase in liver enzymes, which never exceeded three times the upper limit of normal and which in most cases returned to pre-treatment levels before the next dose of DMF. Most cases of elevated liver enzymes have been associated with at least one non-protocol related factor (alcohol consumption, hepatitis and previous anti-HIV therapy with other agents).

Almost all patients showed clinical improvement after 2-3 weeks of treatment. As shown in FIG. 4, each patient exhibited an improved general condition after DMF treatment if the condition was assessed by the Karnov performance scale, according to which the general condition of the patient is determined by a numerical value as follows: 100 = normal, no problems, 90 = capable of performing normal activities, mild symptoms or symptoms of disease, 80 = normal activity and herpetic Other antis exertion, 70 = self-care, unable to perform normal activity or active work, 60 = requires occasional help but is able to cater for most needs, 50 = requires significant help frequent medical care, 40 = incapable, requires special care, 30 = severe incapacity, indicated hospitalization, although there is no immediate risk of death, 20 = very ill, necessary hospitalization and supportive treatment, 10 = moribund, fatal progression progresses rapidly, 0 = dead (see Karnovsky et al., 1984, Cancer 1: 634656). The improvement in neurological symptoms of viral infections was remarkable, microbial therapies for secondary infections were rarely needed. During the first 14 days of DMF treatment, there was a significant reduction in general fatigue and an increase in appetite. All patients increased their weight. Clinical improvement correlated well with disease state as assessed by viral load and CD4 ⁺ cell count. In five out of eight patients, the relative viral load measured by PCR can fit Gompertz curves; this analysis revealed a 88.8% reduction in viral load measured by PCR after 42 days of DMF treatment. In seven out of eight patients, the relative number of CD4 ⁺ T cells may suit Gompertz curves; this analysis showed a 73.4% increase in CD4 ⁺ T-cell counts after 42 days of DMF treatment.

The present invention is not limited to the scope of the disclosed embodiments, which are intended to illustrate particular aspects of the invention. Various other modifications of the invention will be apparent to those skilled in the art from the foregoing descriptions and accompanying drawings. Such modifications are intended to fall within the scope of the appended claims.

All publications cited herein are hereby incorporated by reference in their entirety.

Claims (24)

PATENT CLAIMS A composition for transdermal administration of a therapeutic agent, comprising a reservoir containing or having absorbed therapeutical agent comprising N-hydroxymethyl N-methylformamide (HMMF), N-hydroxymethylformamide (HMF), dihydroxymethylformamide (DHMF), N acetyl S- (N-methylcarbamoyl) cysteine (AMCC), dimethylsulfoxide (DMSO) or pyridine N-oxide. 2. A composition for transdermal administration of a therapeutic agent comprising an external support portion; a permeable membrane suitable for placement against the skin of the animal to be treated; and an adsorbent positioned between the support portion and the permeable membrane, wherein the adsorption agent has an adsorbed therapeutic agent comprised of N, N'-dimethylformamide (DMF), N-hydroxymethyl N-methylformamide (HMMF), N-hydroxymethylformamide (HMF), dihydroxymethylformamide (DHMF) ), N-acetyl, S- (N-methylcarbamoyl) cysteine (AMCC), N-methylformamide (NMF), dimethylsulfoxide (DMSO), formamide, acetamide, methylacetamide, dimethylacetamide, diethyl acetamide, isopropylacetamide, diisopropylacetamide, N-acetyl, N- (β-hydroxyethyl) acetamide, N, N'-di (β-hydroxyethyl) acetamide, N-acetylmorpholine, acrylamide, propionamide, N-fluoromethyl-N-methylformamide, pyridine-N-oxide or amide of formula R ³ - CO-NR ¹ R ² , wherein R ¹ and R ² are independently selected from the group consisting of hydrogen, methyl, halomethyl, saturated or unsaturated C 2 -C 3 alkyl groups and hydroxylated alkyl groups; or R ¹ and R ² are together selected from the group consisting of (CH 2) 4, (CH ₂ ) ₂ and (CH ₂ ) ₂ O (CH ₂ ) ₂ ; and R ³ is selected from the group consisting of hydrogen, methyl, and saturated or unsaturated C 2 -C 3 alkyl groups; a composition for transdermal administration of the therapeutic agent adapted to be placed in contact with the skin of the animal to be treated. A composition for transdermal administration of a therapeutic agent according to claim 2, wherein the therapeutic agent comprises DMF. A composition for transdermal administration of a therapeutic agent according to claim 3, wherein the therapeutic agent comprises at least 0.25 g DMF. A composition for transdermal administration of a therapeutic agent according to claim 4, wherein the therapeutic agent comprises at least 5.0 g of DMF. A composition for transdermal administration of a therapeutic agent according to any one of claims 3 to 5, wherein the therapeutic agent comprises DMF in an amount effective to treat viral or microbial infections or weight loss syndrome and adult humans or children. A composition for transdermal administration of a therapeutic agent according to any one of claims 3 to 6, wherein no other pharmacologically active agent is present in addition to DMF. A composition for transdermal administration of a therapeutic agent according to any one of claims 3 to 7, characterized in that no pharmacologically active agent other than DMF is present that is capable of being systemically absorbed through the skin. A composition for transdermal administration of a therapeutic agent according to any one of claims 3 to 8, wherein the adsorbent is colloidal silica and the permeable membrane is a Teflon ™ membrane with a pore diameter of 0.1 µπι to 0.5 µπι. 10. A method of treating weight loss syndrome, wherein the animal in need of treatment is administered a therapeutic agent comprising N, N'-dimethylformamide (DMF), N-hydroxymethyl N-methylformamide (HMMF), Nhydroxymethylformamide (HMF). , dihydroxymethylformamide (DHMF), N-acetyl, S- (N-methylcarbamoyl) cysteine (AMCC), Nmethylformamide (NMF), dimethylsulfoxide (DMSO), formamide, acetamide, methylacetamide, dimethylacetamide, diethylacetamide, isopropylacetamide, diisopropylacetamide, N- acetylpiperidine, N- (β-hydroxyethyl) acetamide, N, N'-di (β-hydroxyethyl) acetamide, N-acetylmorpholine, acrylamide, propionamide, N-fluoromethyl N-methylformamide, pyridine N-oxide or an amide of formula R ³ -CO-NR ¹ R ² , wherein R ¹ and R ² are independently selected from the group consisting of hydrogen, methyl, halomethyl, saturated or unsaturated C 2 -C 3 alkyl groups and hydroxylated alkyl groups; or R ¹ and R ² are together selected from the group consisting of (CH 2) 4, (CH ₂ ) ₅ and (CH ₂ ) ₂ O (CH ₂ ) 2; and R ³ is selected from the group consisting of hydrogen, methyl, and saturated or unsaturated C 2 -C 3 alkyl groups. The method of claim 10, wherein the therapeutic agent comprises an amide of the formula R ³ -CO-NR ¹ R ² , wherein: R ¹ and R ² are independently selected from the group consisting of hydrogen, methyl, halomethyl, saturated or unsaturated C 2 -C 3 alkyl groups and hydroxylated alkyl groups; or R ¹ and R ² are together selected from the group consisting of (CH ₂ ) ₄ , (CH ₂ ) ₅ and (CH ₂ ) ₂ O (CH ₂ ) ₂ ; and R ³ is selected from the group consisting of hydrogen, methyl, and saturated or unsaturated C ₂ -C 3 alkyl groups. The method of claim 10 or 11, wherein the therapeutic agent comprises an amide of the formula R ³ -CO-NR ¹ R ² wherein: R ¹ and R ² are independently selected from the group consisting of hydrogen, methyl, halomethyl, saturated or unsaturated C ₂ -C 3 alkyl groups and hydroxylated alkyl groups; and R ³ is selected from the group consisting of hydrogen, methyl, and saturated or unsaturated C ₂ -C 3 alkyl groups. The method of any one of claims 10 to 12, wherein the therapeutic agent comprises DMF, HMMF, HMF, DHMF, AMCC, NMF, DMSO, formamide, acetamide, methylacetamide, dimethylacetamide, diethylacetamide, isopropylacetamide, diisopropylacetamide, N-acetylpiperidine. , Ν- (β-hydroxyethyl) acetamide, N, N'-di (β-hydroxyethyl) acetamide, Nacetylmorpholine, acrylamide, propionamide, N-fluoromethyl-N-methylformamide or pyridine-N-oxide. The method of claim 13, wherein the therapeutic agent comprises DMF, HMMF, HMF, DHMF, AMCC, or NMF. The method of any one of claims 10 to 14, further comprising administering at least one additional agent effective in the treatment of HIV infection. 16. The method of claim 15, further comprising administering at least one nucleoside analogue reverse transcriptase inhibitor, at least one non-nucleoside reverse transcriptase inhibitor, or at least one protease inhibitor. The method of any one of claims 14 to 16, wherein the therapeutic agent comprises DMF. The method of any one of claims 13, 15 or 16, wherein the therapeutic agent comprises DMSO. The method of any one of claims 10 to 18, wherein the therapeutic agent is administered transdermally. 20. A method of treating viral or macrobial infections, comprising administering to an animal in need thereof a therapeutic agent comprising N, N'-dimethylformamide (DMF), N-hydroxymethyl N-methylformamide (HMMF), N-hydroxymethylformamide (HMF). , dihydroxymethylformamide (DHMF), N-acetyl, S- (N-methylcarbamoyl) cysteine (AMCC), dimethylsulfoxide (DMSO) or pyridine-N-oxide. 21st The method of claim 20 mark the task s a t ý m that therapeutic substance is given for treatment retro- viral infection. 22nd The method of claim 21 mark the task s a t ý m that retrovirus the infection is HIV infection. The method of any one of claims 20 to 22, further comprising administering at least one additional agent effective in the treatment of HIV infection. The method of any one of claims 20 to 23, further comprising administering at least one nucleoside analogue reverse transcriptase inhibitor, at least one non-nucleoside reverse transcriptase inhibitor, or at least one protease inhibitor. The method of any one of claims 20 to 24, wherein the therapeutic agent is administered transdermally. 26. A vaccine prepared from antibodies derived from the body of animals treated for viral infection according to the method of claim 20.