MXPA98000915A - Use of l-carnitine and derivatives to reduce ceramide levels and for antirretrovi potentiation - Google Patents

Abstract

The present invention relates to a novel use of L-carnitine, the derivatives thereof and their pharmacologically acceptable salts, in combination with antiretroviral drugs to reduce ceramide levels and improve the activity of the aforementioned antiretroviral drugs, for the treatment Therapeutic treatment of HIV infection and SI

Description

USE OF L-CARNITINE AND DERIVATIVES TO REDUCE THE LEVELS OF CERAMIDE AND FOR POTENTIATION ANTIRETROVIRAL DESCRIPTION OF THE INVENTION The present invention relates to a new therapeutic use of L-carnitine, the derivatives thereof, and their pharmacologically acceptable salts in combination with antiretroviral drugs for the therapeutic treatment of HIV infection and AIDS. More particularly, the present invention relates to the use of L-carnitine, to acyl-L-carnitines wherein the acyl group, linear or branched, has from 2 to 6 carbon atoms, and pharmacologically acceptable salts thereof. , in combination with nucleoside-like inhibitors of reverse transcriptase, non-nucleoside inhibitors of reverse transcriptase and inhibitors of HIV protease, to reduce ceramide levels and improve the activity of the aforementioned antiretroviral drugs in infected patients with HIV Most of the pathogenetic mechanisms that contribute to the progression of infection due to REF: 26792 human immunodeficiency virus 1 or 2 (HIV-1, HIV-2) are directly or indirectly related to the general state of activation of the immune system. The chronic activation of the immune system potentiates viral replication via the secretion of a number of cytokines that favor the expression of HIV and by maintaining a reserve of activated immune cells which act as targets or targets for HIV and facilitate its replication. In addition, the state of persistent activation of the immune system induces abnormalities of such nature (eg, increased apoptosis) to lead to a progressive weakening of immune responses. In this way a vicious circle is established: the progressive loss of competence of the immune system - > viral spread - »reduced virus clearance -» chronic activation of the immune system. The above process can last several years until a remarkable deterioration of the immune system occurs, to lead to uncontrolled viral replication and to the onset of opportunistic infections, or to the development of the acquired immunodeficiency syndrome (AIDS). Based on the pathogenetic mechanisms described above, it seems clear that any anti-HIV treatment should be aimed at reducing viral replication and blocking the deterioration of the immune system. As seen in antiretroviral therapy, unfortunately HIV is characterized by a high degree of genetic variability that originates above all in the very substantial lack of precision of reverse transcriptase. The retroviral enzyme lacks enzymatic systems for the control of possible errors in transcription. The result is the emergence of variants of the virus, in a wide range that is a function of viral replication, which are responsible for the progressive avoidance of the immune system and resistance to antiretroviral drugs. In the case of zidovudine (AZT, ZDV) the loss of clinical efficacy in monotherapy situations is a widely recognized fact. Even the most recently discovered antiretroviral agents, for example, zalcitabine [ddc], didanosine [ddl] and lamivudine [3TC] suffer from the same drawback. It has recently been shown that ceramide stimulates the expression of HIV. What is more, ceramide is one of the factors capable of inducing cellular apoptosis, a phenomenon that is increased in subjects with HIV infection and which contributes to the decrease of TCD4 and TCD8 lymphocytes. It seems obvious that changes in the concentration or metabolism of ceramide can affect viral load and cellular apoptosis in HIV-infected subjects (Papp B., et al., AIDS, Res. Hum. Retrovirus, 10 (7) , 775-80). Surprisingly, it has now been found that L-carnitine and derivatives thereof, for example, the acyl-L-carnitines wherein the acyl group, linear or branched has from 2 to 6 carbon atoms and pharmacologically acceptable salts thereof, inhibit the synthesis of ceramide by at least 25%, and when these are used in combination with antiretroviral drugs such as, for example, AZT, stavudine [d4T], fluorotimidine [FLT], azidouridine [Azu], phosphonated acyclic nucleosides [PMEA], specific nucleosides of HIV-1 [TSAO], zalcitabine [ ddC], didanosine [ddl] and lamivudine [3TC], dipyridodiazepinones, tetrahydroimidazobenzo-diazepinones, pyridones or L-drugs, bis-heteroarylpiperazines, alpha-anilinophenyl-acetamide derivatives, quinaxoline derivatives, Ro-31-8959, U-81749 , KNI-227, SC-52151, HOE / BAY 793 and the like, improve the antiretroviral activity and defense of the immune system exerted by these drugs.

The pharmaceutically acceptable salts of L-carnitine or acyl-L-carnitine include, in addition to the internal salts, any salt thereof, with acids which do not give rise to undesirable toxic or collateral effects. These acids are well known to medium-skilled pharmacologists and experts in pharmaceutical technology. Non-limiting examples of suitable salts include, chloride; bromide; I last; aspartate; particularly aspartate acid; citrate, particularly acid citrate; tartrate; phosphate, particularly acid phosphate; fumarate, particularly acid fumarate; glycerophosphate; glucose-phosphate; lactate; maleate, particularly acid maleate; orotato; oxalate, particularly oxalate acid; sulfate, particularly acid sulfate; trichloroacetate; trifluoroacetate and methanesulfonate. Particularly preferred are L-carnitine, acetyl-, propionyl-; butyryl-; valeryl- and isovaleryl-L-carnitine. The combined administration of L-carnitine and its derivatives as defined above plus an antiretroviral agent, is generally conducted via the oral or parenteral routes at daily doses in the range of 1 to 500 mg / kg, with a particular preference for doses of 20 to 100 mg / kg, in a proportion of L-carnitine and its derivatives as defined above, to the antiretroviral agent in the range of 1:40 to 40: 1, with a particular preference for proportions of 1:10 to 10: 1. Conveniently, administration will take the form of a unit dose that includes both active ingredients, and this may also include additional excipients and active ingredients well known to those skilled in the art, such as, for example, dextran, TNF-alpha inhibitors ( for example pentoxifylline), glutathione and other antioxidant drugs (for example, acetylcysteine), immunomodulatory drugs, immunosuppressive or chemotherapeutic agents, vitamins and trace elements. Finally, it should be noted that everything suggests that other basic amino acids, particularly lysine, acylated derivatives of basic amino acids and their pharmaceutically acceptable salts, are capable of reducing ceramide levels and potentiating the activity of antiretroviral drugs for the therapeutic management of HIV infection and AIDS. The purpose of the following examples is to illustrate the invention and these should not be considered in any way as limiting the range of possibilities.

EXAMPLE 1 The effect of administration of a combination of L-carnitine (8 g daily orally for 4 weeks) plus AZT (600 mg daily orally) in 13 patients suffering from AIDS with normal levels of carnitine and acetylcarnitine in serum and intracellularly was evaluated , who had been previously treated with AZT (600 mg daily orally) for at least 6 months. Determinations were made before the combined treatment, while the patients were taking only AZT (T0), after 4 weeks of therapy with the combination of L-carnitine-AZT (TI) and one month after the discontinuation of the treatment with L-carnitine (T2), leaving patients with AZT alone. TCD4 lymphocytes were measured by flow cytometry by means of a specific monoclonal antibody (number of lymphocytes per mm3) and apoptotic lymphocytes by flow cytometry after staining with propidium iodide, quantifying the cells with hypoliploid nuclei (number of lymphocytes) by 50,000 cells). Viral load (number of viral particles per my serum) was determined by quantification of HIV-1 RNA, by means of a polymerase chain reaction (HIV detection system, Amplicor by Roche). The ilcoxon test was used for statistical processing. The results are shown in Table 1 below.

Table 1 T-lymphocytes TCD-J / mm "Lin f oops HIV (apoptotic particles cos / 50,000 viles per mi) cells TO TI T2 TO TI T2 T0 TI 1 195 211 172 85 59 130 3,500 900 2 239 254 264 112 37 84 2,400 1,600 3 172 176 176 90 29 35 2,800 1,600 4 254 287 279 91 40 39 1,700 1,500 141 165 165 59 47 66 62,000 1,100 6 125 205 146 280 81 36 2,200 2,000 7 40 51 47 41 26 88 3,800 3,100 8 309 423 411 82 67 41 5,900 3,900 9 303 502 402 102 20 41 5,300 3,300 47 46 43 65 42 64 2,300 2,300 11 120 120 116 52 52 55 1,400 1,400 Table 1 (continued) Pacieni-e Lin tor-it- - r t-D / mm3 HIV lymphocytes (apoptotic particles / 50,000 viral by my cells TO Ti T2 TO TI T2 TO Ti 12 52 60 41 148 111 104 900 1,300 13 26 378 378 113 38 54 43,000 7,000 Medium 178 221 203 101 50 64 10,553 2,384 Patient TD4 lymphocytes / mm3 HIV lymphocytes (apoptotic particles / 50,000 viral cells per ml) FROM. 101 144 133 61 25 64 19,070 1,661 Significance- 0.001 0.02 0.001 0.04 0.004 cancia Statistical In the same subjects the levels of ceramide in lymphocytes, measured by means of the DAG (diacylglycerol) kinase assay (Cifone M. G. et al., J.

Exp. Med. 180 (4), 1547-52) decreased from 48 ± 8 pmol / 106 lymphocytes measured from combination treatment (T0) to 27 ± 5 pmol / 106 lymphocytes (TI) (P <0.01), rising again to 38 ± 9 pmol / 10D lymphocytes one month after the discontinuation of L-carnitine (T2). These results clearly show that treatment with AZT alone, even when previously prolonged for more than 6 months (T0), does not provide those immunological and virological improvements that are achievable with the combination with L-carnitine-AZT in only 4 weeks (TI ). These improvements tend to decline with the discontinuation of treatment (T2).

EXAMPLE 2 Four subjects with AIDS were treated, with 600 mg of AZT daily orally. Of these, two were also treated with L-carnitine with 3 grams daily. The total duration of treatment was 6 months. Muscle biopsies were performed before and after treatment. The ceramide present at the level of muscle cells was determined before and after treatment, after sonication and genization of the biopsy material. The viral load was determined in the genates of the muscle itself, as described in Example 1. The results are shown in Table 2 below.

Table 2 eramide Ceramide HIV HIV pretreatment after the pretreatment post-treatment (pmol / mg of (mol / mg (particles (protein)) protein) viral / mg of viral / mg of protein protein Patient 1 89 95 3, 800 4, 100 (AZT) Patient 2 95 103 5, 900 5, 800 (AZT) Mean 92 99 4, 850 4, 950 FROM. 4 6 1,485 1,202 mean. s n. Patient statistics 3 129 39 10,200 3,200 (AZT + L- carnitine) Patient 4 79 27 5, 100 1, 300 (AZT + L- carnitine Media 104 33 7, 650 4, 950 FROM. 35 8 3, 606 1,344 mean0.01 0.05 statistical rate It is apparent that treatment with the L-carnitine-AZT combination is distinctly more effective in reducing viral load and ceramide levels, also at the muscular level, compared to treatment with AZT alone.

It is noted that in relation to this date, the best method known by the applicant to carry out the aforementioned invention, is the conventional one for the manufacture of the objects to which it relates.

Having described the invention as above, property is claimed as contained in the following:

Claims (13)

1. The use of L-carnitine, acyl-L-carnitines, characterized in that the acyl group, linear or branched, has from 2 to 6 carbon atoms, and their pharmacologically acceptable salts, for the production of a suitable drug for the reduction of ceramide levels for the therapeutic treatment of HIV infection and AIDS.

2. The use according to claim 1, in combination with nucleoside-like reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors and HIV protease inhibitors, characterized in that they are for the production of a suitable drug to reduce levels of ceramide and the improvement of the activity of the aforementioned antiretroviral drugs for the therapeutic treatment of HIV infection and AIDS.

3. The use according to claim 1 or 2, characterized in that L-carnitine is used.

4. The use according to claim 1 or 2, characterized in that the acyl-carnitine is isovaleryl-L-carnitine.

5. The use according to claim 1, characterized in that a basic amino acid, an acylated basic amino acid or a pharmacologically acceptable salt thereof is used.

6. An orally or parenterally administrable pharmaceutical composition, for reducing ceramide levels, for the therapeutic treatment of HIV infection and AIDS, characterized in that the composition comprises as an active ingredient an amount of L-carnitine, acyl-L-carnitines wherein the acyl group, linear or branched, has from 2 to 6 carbon atoms and the pharmacologically acceptable salts thereof, effective to reduce the levels of ceramide, and at least one pharmacologically acceptable excipient.

7. A pharmaceutical composition, orally or parenterally administrable for the therapeutic treatment of HIV infection and AIDS, in combination with the administration of nucleoside reverse transcriptase inhibitors, non-nucleoside inhibitors of reverse transcriptase inhibitors or protease inhibitors of the HIV, characterized the composition because it comprises an amount of L-carnitine, acyl-L-carnitines wherein the acyl group, linear or branched, has from 2 to 6 carbon atoms and the pharmacologically acceptable salts thereof, effective to reduce the ceramide levels and viral load, to improve the immunocompetence of HIV-infected patients, and at least one pharmacologically acceptable excipient.

8. An oral or parenterally administrable pharmaceutical composition in unit dose form, for reducing ceramide levels and improving the efficiency of antiretroviral drugs for the therapeutic treatment of HIV infection and AIDS, characterized in that the composition comprises at least one antiretroviral drug selected from nucleoside-like inhibitors of reverse transcriptase, non-nucleoside inhibitors of reverse transcriptase or inhibitors of HIV protease, and an amount of L-carnitine, acyl-L-carnitines wherein the acyl group, linear or branched, it has from 2 to 6 carbon atoms and pharmacologically acceptable salts thereof, effective to reduce ceramide levels and viral load, and improve immunocompetence of HIV infected subjects, and at least one pharmacologically acceptable excipient.

9. The pharmaceutical composition according to claim 6 or 7, characterized in that the amount of L-carnitine, acyl L-carnitines wherein the acyl group, linear or branched having from 2 to 6 carbon atoms and the pharmacologically acceptable salts of the same, is such that the daily dose of L-carnitine, acyl-L-carnitines wherein the acyl group, linear or branched, has from 2 to 6 carbon atoms and the pharmacologically acceptable salts thereof and the antiretroviral drugs, is from 1 to 500 mg / kg, and the ratio between L-carnitine and acyl-L-carnitines wherein the acyl group, linear or branched has from 2 to 6 carbon atoms, and its pharmacologically acceptable salts, and the drug antiretroviral, is 1:40 to 40: 1.

10. The pharmaceutical composition according to claim 6 or 7, characterized in that the amount of L-carnitine, acyl L-carnitines wherein the linear or branched acyl group has from 2 to 6 carbon atoms and their pharmacologically acceptable salts, is such that the daily dose of L-carnitine, acyl-L-carnitines wherein the linear or branched acyl group has from 2 to 6 carbon atoms and the pharmacologically acceptable salts thereof and the antiretroviral drug, is from 20 to 100 mg / kg , and the ratio between L-carnitine, acyl-L-carnitines wherein the acyl group, linear or branched has from 2 to 6 carbon atoms and pharmacologically acceptable salts thereof, and the antiretroviral drug is from 1:10 to 10: 1

11. The orally or parenterally administrable pharmaceutical composition according to claim 8, in unit dose form, characterized in that the amount of L-carnitine, acyl-L-carnitines wherein the linear or branched acyl group has from 2 to 6 carbon atoms. carbon and its pharmacologically acceptable salts, is such that the daily dose of L-carnitine, acyl-L-carnitines wherein the linear or branched acyl group has from 2 to 6 carbon atoms and the pharmacologically acceptable salts thereof and the antiretroviral drug , is from 1 to 500 mg / kg, and the ratio between L-carnitine, acyl-L-carnitines wherein the acyl group, linear or branched has from 2 to 6 carbon atoms and the pharmacologically acceptable salts thereof, and the antiretroviral drug is 1:40 to 40: 1.

12. The orally or parenterally administrable pharmaceutical composition according to claim 8, in unit dose form, characterized in that the amount of L-carnitine, acyl-L-carnitines wherein the linear or branched acyl group has from 2 to 6 carbon atoms. carbon and its pharmacologically acceptable salts, is such that the daily dose of L-carnitine, acyl-L-carnitines wherein the linear or branched acyl group has from 2 to 6 carbon atoms and the pharmacologically acceptable salts thereof and the antiretroviral drug , is from 20 to 100 mg / kg, and the ratio between L-carnitine, acyl-L-carnitines wherein the acyl group, linear or branched has from 2 to 6 carbon atoms and the pharmacologically acceptable salts thereof, and the antiretroviral drug is from 1:10 to 10: 1.

13. The composition according to any one of claims 1 to 12, characterized in that it also comprises at least one component selected from dextran, alpha-TNF inhibitors, antioxidant drugs, immunomodulators, immunosuppressants, chemotherapeutic agents, vitamins and trace elements.