Title:  Combination therapy for HIV infections

United States Patent:  6,696,422

Issued:  February 24, 2004

Inventors:  Hirschman; Shalom Z. (Riverdale, NY)

Assignee:  Advanced Viral Research Corp. (Hallandale, FL)

Appl. No.:  316374

Filed:  May 21, 1999

Abstract

The present invention discloses a method of treating patients having AIDS or HIV infections by parenterally administering Product R, a peptide-nucleic acid preparation, in a combination with one or more antiviral agents useful for treating AIDS or HIV infections including HIV protease inhibitors and nucleoside analogs.

SUMMARY OF THE INVENTION

Accordingly, an object of this invention is to provide a method for treating a patient having AIDS or HIV infections by administering an effective treatment amount of Product R in combination with one or more antiviral agents useful for treating AIDS or HIV infections, such as nucleoside analogs, to the patient.

Another object of the present invention is to provide a method for treating a patient having AIDS or HIV infections by a combination of an effective treatment amount of Product R and one or more nucleotide analogs and/or HIV protease inhibitors.

According to the present invention, the identified patient is treated by administering parenterally an effective treatment amount of Product R from about 5 microliters to about 40 microliters per kilogram of body weight per day in a sterile injectable formulation.

Other objects and features of the present invention will become apparent from the following detailed description considered in conjunction with the accompanying drawings. It is to be understood, however, that the drawings are designed solely for purposes of illustration and not as a definition of the limits of the invention, for which reference should be made to the appended claims.

DETAILED DESCRIPTION OF THE PRESENTLY PREFERRED EMBODIMENTS

As used herein, Product R is the product produced according to either of the following methods.

Method I For Preparing Product R

Suspend about 35.0 g of casein, about 17.1 g of beef peptone, about 22.0 g of nucleic acid (RNA), about 3.25 g bovine serum albumin in about 2.5 liters of water for injection USP at about 3 to 7^oC. in a suitable container and gently stir until all the ingredients have been properly wet. Carefully add while stirring about 16.5 g of sodium hydroxide (reagent grade ACS) and continue stirring until sodium hydroxide completely dissolved. Autoclave at about 9 lbs pressure and 200-230^oF. for a period of time until RNA is completely digested, for example, about 4 hours. At the end of the period, the autoclave is stopped and the reaction flask and contents are permitted to slowly cool to ambient temperature. Then cool for at least six hours at about 3-8^oC. The resulting solution is filtered through 2 micron and 0.45 micron filters using inert gas such as nitrogen or argon at low pressure (1-6 psi). In a similar manner the solution is filtered again through 0.2 micron pyrogen retention filters. The resulting filtrate is sampled and assayed for total nitrogen. A calculation is then performed to determine the quantity of cooled water for injection to be added to the filtrate to yield a diluted filtrate with a nitrogen content between about 165-210 mg/ml, the final volume is approximately 5 liters. The pH is then adjusted with either concentrated HCl (reagent grade ACS) or 1.0 normal NaOH to about 7.3-7.6 range. The diluted solution is then filtered again through 0.2 micron filters with inert gas at low pressure. The final filtrate is then filled and sealed into 2 ml glass ampules while in an inert gas atmosphere. The ampules are collected and autoclave for final sterilization at 240^oF. and 20 to 30 pounds pressure for about 30 minutes. Following the sterilization cycle, the ampules with Product R are cooled and washed.

All quantities are subject to plus or minus 2.5% variation for pH, volume, and analytical adjustments.

Method II For Preparing Product R

Suspend about 35.0 g of casein, about 17.1 g of beef peptone, about 22.0 g of nucleic acid (RNA), about 3.25 g bovine serum albumin in about 2.5 liter of water for injection USP at about 3 to 7^oC. in a suitable container and gently stir until all the ingredients have been properly wet. Slowly add while stirring about 11.75 ml of hydrochloric acid (reagent grade ACS) and continue stirring until hydrochloric acid is completely dissolved. Autoclave at about 9 lbs pressure and 200-230^oF. for a period of time until RNA is completely digested, for example, about 4 hours. At the end of the period, the autoclave is stopped and the reaction flask and contents are permitted to slowly cool to ambient temperature. Then cool for at least six hours at about 3-8^oC. The resulting solution is filtered through 2 micron and 0.45 micron filters using inert gas such as nitrogen or argon at low pressure (1-6 psi). In a similar manner the solution is filtered again through 0.2 micron pyrogen retention filters. The resulting filtrate is sampled and assayed for total nitrogen. A calculation is then performed to determine the quantity of cooled water for injection to be added to the filtrate to yield a diluted filtrate with a nitrogen content between about 165-210 mg/ml, the final volume is approximately 5 liters. The pH is then adjusted with either concentrated HCL (reagent grade ACS) or 35% (w/v) of NaOH to about 7.3-7.6 range. The diluted solution is then filtered again through 0.2 micron filters with inert gas at low pressure. The final filtrate is then filled and sealed into 2 ml glass ampules while in an inert gas atmosphere. The ampules are collected and autoclave for final sterilization at 240^oF. and 20 to 30 pounds pressure for about 30 minutes. Following the sterilization cycle, the ampules with Product R are cooled and washed.

All quantities are subject to plus or minus 2.5% variation for pH, volume, and analytical adjustments.

HIV protease inhibitors include oligopeptide analogs, such as saquinavir (Roche Laboratories), indinavir (Merck) or ritonavir (Abbott Laboratories), which are fully described in detail in U.S. Pat. Nos. 5,413,999 and 5,476,874. The contents of which are hereby incorporated by reference in their entirety.

The antiviral agents useful for a combination therapy of AIDS or HIV infections other than HIV protease inhibitors is listed in Table I.

It will be understood that the scope of combinations of Product R with antiviral agents is not limited to the list in the above Table I or described above, but includes in principle any combination with any pharmaceutical composition useful for treating patients having AIDS or HIV infections.

Preferred combinations are concurrent or alternating treatments of one or more HIV protease inhibitors or antiviral agents useful for treating AIDS or HIV in forms of nucleoside analogs, such as AZT, ddI or ddC, and Product R. Preferably, the HIV protease inhibitors or antiviral agents useful for treating AIDS or HIV infections are administered first, e.g. for one week, to quickly decrease the viral load; Product R then is administered to stimulate the immune system to eradicate the HIV infections. For patients who are very ill with opportunistic infections, Product R may be administered to stimulate the immune system first before and then after the indicated antiviral agents are applied.

Alternatively, two antiviral agents useful for treating AIDS or HIV such as lamivudine and AZT, together with one protease inhibitor such as ritonavir, may be administered concurrently first until the viral load is no longer measurable, then Product R is administered to eradicate the HIV infections.

Or, Product R may be administered concurrently with two antiviral agents such as lamivudine and nevirapone, one protease inhibitor such as indinavir.

For the patient having HIV infections, whether exhibiting AIDS symptoms or having antibodies against HIV or having asymptomatic infections, a suitable effective dose of Product R may be in the range of from about 5 microliters to about 40 microliters per kilogram of body weight per day, preferably in the range of about 10 microliters to about 25 microliters per kilogram of body weight per day. Most preferably Product R is administered in an amount of about 30 microliters per kilogram of body weight per day for about one week, followed by about 15 microliters per kilogram of body weight per day in a sterile injectable formulation. The desired dose may be administered as two, three or more sub-doses at appropriate intervals, generally equally spread in time throughout the day. Preferably, the full daily dose is administered in one administration.

The dose of the antiviral agents or HIV protease inhibitors to be co-administered with Product R can be readily determined by those skilled in the art, based on the usual patient symptoms, and severity of the diseases.

Product R may be administered by any suitable injection route including, but not limited to, intravenously, intraperitoneally, subcutaneously, intramuscularly, and intradermally, etc. The presently preferred route of administration is intramuscularly. It will be appreciated that the preferred route may vary with, for example, the condition and age of the recipient.

While it is possible for Product R to be administered as part of a pharmaceutical formulation, it is preferable to present it alone, although it may be administered at about the same time as one or more other pharmaceuticals are independently administered. If Product R is administered as part of a pharmaceutical formulation, the formulations of the present invention comprise at least one administered ingredient, as above defined, together with one or more acceptable carriers thereof and optionally other therapeutic ingredients. The carrier(s) must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

The formulations may conveniently be presented in unit-dose or multi-dose containers, e.g. sealed ampules and vials.

Thus, while there have been shown and described and pointed out fundamental novel features of the invention as applied to preferred embodiments thereof, it will be understood that various omissions and substitutions and changes in the form and details of the method illustrated, and in their operation, may be made by those skilled in the art without departing from the spirit of the invention. For example, it is expressly intended that all combinations of those elements and/or method steps which perform substantially the same function in substantially the same way to achieve the same results are within the scope of the invention. It is the intention, therefore, to be limited only as indicated by the scope of the claims appended hereto.

Claim 1 of 6 Claims

I claim:

1. A method for reducing human immunodeficiency viral load while increasing CD4 cell count in a patient having acquired immunodeficiency syndrome (AIDS) resulting from human immunodeficiency virus (HIV) infection, comprising the steps of:

(a) administering to said patient an effective HIV infection treatment mount of a Product R and an effective HIV infection treatment amount of at least one other antiviral agent useful for treating HIV infection comprising at least one nucleoside analog; and

(b) discontinuing said administration of said one other antiviral agent and continuing to administer said effective HIV infection treatment amount of a Product R to said patient following step (a);

wherein said Product R is produced according to Method I, and wherein the step of using about 16.5 g of sodium hydroxide for preparing said Product R is employed.

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