CA2085367A1 - Hiv-transactivators to treat aids - Google Patents
Hiv-transactivators to treat aidsInfo
- Publication number
- CA2085367A1 CA2085367A1 CA002085367A CA2085367A CA2085367A1 CA 2085367 A1 CA2085367 A1 CA 2085367A1 CA 002085367 A CA002085367 A CA 002085367A CA 2085367 A CA2085367 A CA 2085367A CA 2085367 A1 CA2085367 A1 CA 2085367A1
- Authority
- CA
- Canada
- Prior art keywords
- human
- neomycin
- poly
- hydroxybenzyl
- hiv
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
- A61P31/18—Antivirals for RNA viruses for HIV
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/22—Cyclohexane rings, substituted by nitrogen atoms
- C07H15/222—Cyclohexane rings substituted by at least two nitrogen atoms
- C07H15/226—Cyclohexane rings substituted by at least two nitrogen atoms with at least two saccharide radicals directly attached to the cyclohexane rings
- C07H15/228—Cyclohexane rings substituted by at least two nitrogen atoms with at least two saccharide radicals directly attached to the cyclohexane rings attached to adjacent ring-carbon atoms of the cyclohexane rings
- C07H15/232—Cyclohexane rings substituted by at least two nitrogen atoms with at least two saccharide radicals directly attached to the cyclohexane rings attached to adjacent ring-carbon atoms of the cyclohexane rings with at least three saccharide radicals in the molecule, e.g. lividomycin, neomycin, paromomycin
Abstract
N-poly-(o-hydroxybenzyl) Neomycin B or salts and/or hydrates thereof, can be used to treat humans infected with one or more than one strain of a human immunodeficiency virus.
Description
WO92/02530 7~ ) 3~ PCI`/US91/04849 i ~exa-l~l-(o-hy~roxybenzyl)Neomycir, B for inhibiting human re~roviruses and ror ~he treatmenl or AI~S
FIELD OF THE INVENTION
This invention relates to N-po1y-(o-hydroxybenzyl) Neomycin B or salts and/or hydrates thereof, as a drug to treat a human cell system, including a human patient, possibly infected with S a human retrovirus (HRV) to prevent or retard the -further repiication of the HRV in that human system or patient.
BACKGROUND OF THE INVENTION
An estimated one to one and one-half million people in the United States are infected with a human retrovirus, the human immunodeficiency virus type I, HIV-I, which is the etiologicial ageDt of acquired immunodeficiency syndrome, AIDS, Norman, C., Science, 661 662 (1986). Of those infected, an estimated two hundred and fifty thousands people will develop AIDS in the ne~t five years, Curran, J.W., et al., Science, 1352-1357 (1985). Oo March 20, 1987, the FDA
approved the use of the compound, zidovudine (AZT), to treat AIDS patients with a recent initial episode of pneumocystis carinii pneumonia, AIDS patients with conditions other than pneumocystis carinii pneumonia or patients infected with the ViNS Wi~l an absolute CD4 Iymphocyte count of less than 200/m n3 in the peripheral blood. AZT is a known inhibitor of viral reverse transcript-ase, an enzyme necessary for human immunodeficiency virus replication.
U.S. Patent 4,724,232 claims a method of treating humans having acquired immunodeficiency syndrome utilizing 3'-azido-3'~eoxy-thymidine (azidothymidine, AZT).
European Patent Application 88306456.0 (Publication No. 0 0300 687; 2S.01.89 Bulletin 89/04) describes a method of inhibiting human imrnunodeficiency virus utilizing a therapeutically effective amount of an antiviral agent to attack the first splice acceptor site of tat nI gene of HIV.
Oligodeoxyyribonucleoside methylphosphonates (OMPs) are disclosed as useful antiviral agents for the treatment of HIV.
Neomycin B free base is well known and commercially availabl~ from various sources, including for example Sigma (Sigma #N 1876); February 1986, page 869.
SUMMARY OF THE INVENTION
This invention is a method for treating a human infected with one or more than one strain of a human immunodeficiency virus which comprises administering an effective arnount of a N-poly-(o-hydroxybenzyl) Neomycin B, or salts and/or hydrates thereof, to the infected human.
Examples of alkali metal salt forms of N-poly-(o-hydroxybenzyl) Neomycin B
include the sodium, potassium and lithium salts thereof.
DETAILED DESCRlPTION OF THE INVENTION
The term human retrovirus (HRV) includes human immunodeficiency virus type 1, human immunodeficiency virus type II, or strains thereof, as we}l as human T cell leukemia virus 1 and 2 (Hll~Y-l and HTLV-2) strains apparent to one skilled in the art, which belong to the same viral WO 92/02530 PCl/US91/04849 "J3~ -2-families and which create similar physiological effects in humans as various human retroviruses.
Patients to be treated would be those individuals: I) infected with one or more than one strain of a human retrovirus as detennined by the presence of either measurable viral antibody or antigen in the serum and 2) having either a symptomatic AIDS defining infection such as i) 5 disseminated histoplasmosis, ii) isopsoriasis, iii) bronchial and pulmonary candidiasis including pneumocystic pneumonia iv) non-Hodgkin's Iymphoma or v) Kaposi's sarcoma and being less than si~ty years old; or having an absolute CD4 Iymphocyte count of less than 200/mrn3 in the peripheral blood. Treatment would consist of maintaining an isnhibitory level of the compound used according to this invention in the patient at all times and would continue until the occurrence of 10 a second symptomatic AIDS dehning infection indicates alternate therapy is needed.
The structural forsnula of N-poly-(o-hydroxyben~yl) Neomycin B is illustrated in Chart 1, where R is -(2-hydro%y)phenyl.
This compound as depicted in the structure charts as the free base, however phar-macologically acceptable salts and/or hydrates thereof, can be used and administered in practicing 15 the method claimed in this invention. Pharmacologically acceptable salts refers to those salts of the compounds claimed in this invention which would be readily apparent to a manufacturing pharmaceutical chemist to be equivalent to the parent compound in properties such as formulation, stabiliey, patient acceptance and bio availability.
Those skilled in the art would know how to formulate the compounds used to practice the 20 method claimed in this invention into appropriate pharmaceutical dosage for ns. Examples of the dosage forms include oral formulations, such as tablets or capsules, or parenteral formulations, such as sterile solutions.
When the compound used to practice the method claimed in this invention are administered orally, an effective amount is from about 1 to 100 mg per kg per day. A typical unit dose for a 25 iO kg human would be from about 50 mg to 1000 mg, preferably 200 mg to 1000 mg taken one to four times per day. Either solid or fluid dosage forms can be prepared for oral administration.
Solid compositions are prepared by mixing the compounds used to practice the method claimed in this invention with conventional ingredients such as talc, magnesium stearate, dicalcium phosphate, magnesium aluminum silicate, calcium sulfate, starch, lactose, acacia, methyl cellulose, or 30 functionally similar pharmaceutical diluents and carriers. Capsules are prepared by mixing ~e compounds used to practice the method claimed in this invention with an inert phannaceutical diluent and placing the mixture into an appropriately sized hard gelatin capsule. Soft gela~in capsules are prepared by machine encapsulation of a slurry of the compounds used to practice the me~od claimed in this invention with an acceptable inert oil such as vegetahle oil or light liquid 35 petrolatum. SYNPS are prepared by dissolving ~te compounds used to practice the method claimed in this invention in an aqueous vehicle and adding sugar, aromatic flavoring agents and preserva-WO 92/02530 2 ~ 8 3 6 7 Pcr/us91/04849 . .
FIELD OF THE INVENTION
This invention relates to N-po1y-(o-hydroxybenzyl) Neomycin B or salts and/or hydrates thereof, as a drug to treat a human cell system, including a human patient, possibly infected with S a human retrovirus (HRV) to prevent or retard the -further repiication of the HRV in that human system or patient.
BACKGROUND OF THE INVENTION
An estimated one to one and one-half million people in the United States are infected with a human retrovirus, the human immunodeficiency virus type I, HIV-I, which is the etiologicial ageDt of acquired immunodeficiency syndrome, AIDS, Norman, C., Science, 661 662 (1986). Of those infected, an estimated two hundred and fifty thousands people will develop AIDS in the ne~t five years, Curran, J.W., et al., Science, 1352-1357 (1985). Oo March 20, 1987, the FDA
approved the use of the compound, zidovudine (AZT), to treat AIDS patients with a recent initial episode of pneumocystis carinii pneumonia, AIDS patients with conditions other than pneumocystis carinii pneumonia or patients infected with the ViNS Wi~l an absolute CD4 Iymphocyte count of less than 200/m n3 in the peripheral blood. AZT is a known inhibitor of viral reverse transcript-ase, an enzyme necessary for human immunodeficiency virus replication.
U.S. Patent 4,724,232 claims a method of treating humans having acquired immunodeficiency syndrome utilizing 3'-azido-3'~eoxy-thymidine (azidothymidine, AZT).
European Patent Application 88306456.0 (Publication No. 0 0300 687; 2S.01.89 Bulletin 89/04) describes a method of inhibiting human imrnunodeficiency virus utilizing a therapeutically effective amount of an antiviral agent to attack the first splice acceptor site of tat nI gene of HIV.
Oligodeoxyyribonucleoside methylphosphonates (OMPs) are disclosed as useful antiviral agents for the treatment of HIV.
Neomycin B free base is well known and commercially availabl~ from various sources, including for example Sigma (Sigma #N 1876); February 1986, page 869.
SUMMARY OF THE INVENTION
This invention is a method for treating a human infected with one or more than one strain of a human immunodeficiency virus which comprises administering an effective arnount of a N-poly-(o-hydroxybenzyl) Neomycin B, or salts and/or hydrates thereof, to the infected human.
Examples of alkali metal salt forms of N-poly-(o-hydroxybenzyl) Neomycin B
include the sodium, potassium and lithium salts thereof.
DETAILED DESCRlPTION OF THE INVENTION
The term human retrovirus (HRV) includes human immunodeficiency virus type 1, human immunodeficiency virus type II, or strains thereof, as we}l as human T cell leukemia virus 1 and 2 (Hll~Y-l and HTLV-2) strains apparent to one skilled in the art, which belong to the same viral WO 92/02530 PCl/US91/04849 "J3~ -2-families and which create similar physiological effects in humans as various human retroviruses.
Patients to be treated would be those individuals: I) infected with one or more than one strain of a human retrovirus as detennined by the presence of either measurable viral antibody or antigen in the serum and 2) having either a symptomatic AIDS defining infection such as i) 5 disseminated histoplasmosis, ii) isopsoriasis, iii) bronchial and pulmonary candidiasis including pneumocystic pneumonia iv) non-Hodgkin's Iymphoma or v) Kaposi's sarcoma and being less than si~ty years old; or having an absolute CD4 Iymphocyte count of less than 200/mrn3 in the peripheral blood. Treatment would consist of maintaining an isnhibitory level of the compound used according to this invention in the patient at all times and would continue until the occurrence of 10 a second symptomatic AIDS dehning infection indicates alternate therapy is needed.
The structural forsnula of N-poly-(o-hydroxyben~yl) Neomycin B is illustrated in Chart 1, where R is -(2-hydro%y)phenyl.
This compound as depicted in the structure charts as the free base, however phar-macologically acceptable salts and/or hydrates thereof, can be used and administered in practicing 15 the method claimed in this invention. Pharmacologically acceptable salts refers to those salts of the compounds claimed in this invention which would be readily apparent to a manufacturing pharmaceutical chemist to be equivalent to the parent compound in properties such as formulation, stabiliey, patient acceptance and bio availability.
Those skilled in the art would know how to formulate the compounds used to practice the 20 method claimed in this invention into appropriate pharmaceutical dosage for ns. Examples of the dosage forms include oral formulations, such as tablets or capsules, or parenteral formulations, such as sterile solutions.
When the compound used to practice the method claimed in this invention are administered orally, an effective amount is from about 1 to 100 mg per kg per day. A typical unit dose for a 25 iO kg human would be from about 50 mg to 1000 mg, preferably 200 mg to 1000 mg taken one to four times per day. Either solid or fluid dosage forms can be prepared for oral administration.
Solid compositions are prepared by mixing the compounds used to practice the method claimed in this invention with conventional ingredients such as talc, magnesium stearate, dicalcium phosphate, magnesium aluminum silicate, calcium sulfate, starch, lactose, acacia, methyl cellulose, or 30 functionally similar pharmaceutical diluents and carriers. Capsules are prepared by mixing ~e compounds used to practice the method claimed in this invention with an inert phannaceutical diluent and placing the mixture into an appropriately sized hard gelatin capsule. Soft gela~in capsules are prepared by machine encapsulation of a slurry of the compounds used to practice the me~od claimed in this invention with an acceptable inert oil such as vegetahle oil or light liquid 35 petrolatum. SYNPS are prepared by dissolving ~te compounds used to practice the method claimed in this invention in an aqueous vehicle and adding sugar, aromatic flavoring agents and preserva-WO 92/02530 2 ~ 8 3 6 7 Pcr/us91/04849 . .
tives. Elixirs are prepared using a hydroalcoholic vehicle such as ethanol, suitable sweeteners such as sugar or saccharin and an aromatic flavoring agent. Suspensions are prepared with an aqueous vehicle and a suspending agent such as acacia, tragacanth, or methyl celhliose.
When the compound used to practice the method claimed in this invention are administered 5 parenterally, it can be given by injection or by intravenous infusion. An effective amount is from about 1 to 100 mg per kg per day. Parenteral solutions are prepared by dissolving ~e compounds used to practice the method claimed in this invention in water and filter sterilizing the solution before placing in a suitable sealable vial or arnpule. Parenteral suspensions are prepared in substantia}ly the same way except a sterile suspension vehicle is used and the compounds used to 10 practice the method claimed in this invention are sterilized wi~h ethylene oxide or suitable gas before it is suspended in the vehicle.
The exact route of administration, dose, or frequency of administration would be readily determined by those skill in the art and is dependant on the age, weight, general physical condition, or other clinical symptoms specific to the patient to be treated.
Patients to be treated would be those individuals: I) infected with one or more than one strain of a human immunodeficiency virus as determined by the presence of either measurable viral antibody or antigen in the serum and 2) having either a symptomatic AIDS defining infection such as i) disseminated histoplasmosis, ii) isoporiasis, iii) bronchial and pulmonary candidiasis including pneumocystis pneumonia, iv) non-Hodgkin's Iymphoma, or v) Kaposi's sarcoma and being less than sixty years old; or having an absolute CD4 Iymphocyte count of less than 200/mm3 in the peripheral blood. Treatment would consist of maintaining an inhibitory level of the compounds of this invention in the patient at all times and would continue until the occurrence of a second symptomatic AIDS defining infection indicates alternate therapy is needed.
Preparation 1: Preparation of N-poly-(o-hydroxybenzyl) Neomycin B
Part A:
12.33 g. ~20mM) of Neomycin B free base is dissolved in 25 rnl. of water. The solution is diluted with 50 ml of methanol. The resulting solution is mixed with an agueous methanol solution of salicylaldehyde (SAL; salicylic aldehyde; 2-hydroxybenzaldehyde) consisting of 17.1 g. (140 mM) of the SAL, 100 ml methanol and 200 ml water).
The resulting solution is heated to boiling, then allowed to stand at room temperature for 48 hours. The mixture is heated to boiling and 250 ml of hot water added and ~he mixture refrigerated at 0-5 C. for 24 hours. The supernatant is decanted and the resinous product dissolved in the methanol:dioxane (80:20), slurried with 2 g. of DARCO G60 charcoai and filtered though diatomaceous earth (Celite). The filtrate is added with stirring to 300 ml of ether to precipitate the desired Neomycin B Schiff base. The mixture is refrigerated overnight, and the supernatant decanted and the solid dried in vacuo.
WO 92/~2530 P(~/US9l/04849 3~ ~-Part B:
The product of Part A is dissolved in 200 ml methanol and hydrogenated at room temperaNre at 50 psi initial pressure for 24 hours with 1.0 g of PtO2 (Adarns' catalyst). The pressure drop is 7.0 psi per 100 mM. The product is fil~ered through diatomaceous earth (Celite) S and the filtrate concentrated 1n vacuo ~o about 50 ml and resulting concentra~e added slowly to 300 ml of ether. The resulting mixture is refrigerated overnight, fill~ered/decanted and the solid product dried initially at room temperan~re in vacuo for several days and finally overnight at 50 C. in a ~acuum oven to yield 9.5 g of N-poly-(o-hydroxybenzyl) Neomycin B. Analytical resul~s: N, 6.~8;
eq. wt. 216.1; Molecular Weight (MW) 1296.6; ~R 3260, 1605, 1586, 1485, 1045, 1030, 750.
Without further elaboration, those skilled in the art can practice the present invention to its fullest extent. The following detailed examples further describe how to use ~he compounds claimed in this invention to treat humans infected with one or more than one strain of a human immunodeficiency virus. These exdmpl~s are merely illustrative and are not limitations of the preceding disclosure. Those skilled in the art will promptly recognize appropriate Yariations from 15 the examples. In each exarnple, any compound claimed in this invention could replace the compound used in the particular example.
Example 1 Hard Gelatin Capsules One thousand two-piece hard gelatin capsules for oral use, each capsule containing 50 mg of N-poly-(o-hydroxybenzyl) Neomycin B, are prepared from the following:
N-poly-(o-hydroxybenzyl) ~leomycin B 50 grn Lactose 100 gm Cornstarch 20 gm Talc 20 gm Magnesium Stearate 2 gm The N-poly-(o-hydroxybenzyl) Neomycin B is added to the o~her ingredients, mixed and encapsulated in the usual manner.
Example 2 Tablets One thousand tablets, each containing 50 mg of N-poly-(o-hydroxybenzyl) Neomycin B are prepared f~om the following:
N-poly-(o-hydroxybenzyl) Neomycin B 50gm Lactose 75 gm Comstarch 50 gm Magnesium Stearate 4 gm I:ight liquid petrolatum S gm ;, ' ' ' ':
' wo92/û2530 2 0 8 5 3 6 7 Pcr/US9l/04849 The N-poly-(o-hydroxybenzyl) Neomycin B is added to the other ingredients, mi%ed and slugged. The slugs are broken down by forcing through a number sixteen screen. The resulting granules are then compressed into tablets.
Example 3 Parenteral solution A sterile aqueous solution for parenteral intravenous injection containing 150 mg of 1-(~
chlorobenzoyl)-N-poly-(o-hydroxybenzyl) Neomycin B in one liter of solution is prepared from the following:
N-poly-(o-hydroxyben~yl) Neomycin B 150 mg Water for injection, qs 1000 mg The N-poly-(o-hydroxybenzyl) Neomycin B is sterilized, added to the sterile water, filled into sterile containers and sealed.
The utility of this invention is demonstrated by the ability of N-poly-(o-hydroxyben7yl) Neomycin B to practice the method claimed in this invention to inhibit HIV-I tat media~ed transactivation, enzyme essentia~ for human immunodeficiency virus replication. HIV contains various genes, including ntatn which encodes a trans-activator (TAT,\ that functions in the infected cell by increasing the levels of steady-state viral mRNA as well as the transl$ional ~r~ilization of this mRNA. TAT ;s both essential for HIV replication and not structurally related to any norrnal cellular protein. nA Rapid, Quantitative Bioassay Based on the Human Immunodeficiency Virus Trans-Activatorn is described in AIDS RESEARCH AND HUMAN RETROVIRUSES, Vol. 5, Number 5, 1989, pp 507-516. The results (% Inhibition in sCD4 levels) in Table 1.
The anti-HlV utility of this invention is further demonstrated by determining the levels of HIV p24 and HIV RNA in culture supernatants of human peripheral blood Iymphocytes (PBLs) 3 and 4 days after HIV infection. The HIV infectivity experiments were conducted in primary cultures of (PBLs) with HTLV-llIB. Cultures were infected in triplicate with HIV, Compound I
Z5 added, and 3 to 4 days after infection the levels of HIV p24 synthesized and released were quantified by an enzyme-linked immunosorbent assay (ELISA) ~R. Maiolini and R. Masseyeff, 1.
Irnmunol. Methods 8, 223 (1975)] using a monoclonal capnlre antibody to HIV p24 (DuPont, Wilmington, DE) and recombinant HIV p24 calibration standard (MicroGenSys, West Haven, CI ).
The a nounts of HIV RNA synthesized in these infected cultures were also determined by hybridization analysis with a HlV-specific -32P labeled probe. The absolute HIV RNA levels were determined by norrnalizing hybridization values to values obtained from a standard preparation of HIV RNA hybridization .n pa.allel. The results are reported in Table 11.
The proliferation of mitogen-stimul3ted PBLs 4 days after exposure to Compound I was quantified relative to nondrug-treated controls to ensure that the infectivity experiments were conducted a~ non-cytotoxic concentrations of drug. At 40 ug/ml of Compound 1 did not inhibit PBL proliferation. The results are reported in Table In.
.
WO 92t02530 PCr/US91/04849 ~, ~
CHART I
5 Compound No. l N CH R
N0~ C~2 CN2 R~CH2NH _~ NH
,~
HO-CH2 o HO
R = 43 H
' WO 92/02530 PCI/IJ~;91/~4849 ' -7- 2V~367 TABLE I
INHIBITION OF HIY-1 tat MEDIATED TRANSACTIVATION
, ~;
uglml sCD4 in Culture Supernatant % Inhibition (ug/ml) in sCD4 Levels . 10 20 6.48 83 22. 15 43 .~ .
43.8 0 . .
0 38.7 o .
.
, "
.
. . ~
S i TABLE Il Inhibition of HIV-I Replication of U~466 % Inhibition U 6466 ~Mg/rnl) p24 HIV RNA
Day Day _ 4 86 8 l 82 66 0.4 22 0 12 9 0.04 20 4 23 17 ... .
.
:
WO 92/02530 PCr/US91/04849 9 2~3~7 U 6466 (ug/ml)% Cell Viability 0.4 g5 0.04 g6 .
~. :
When the compound used to practice the method claimed in this invention are administered 5 parenterally, it can be given by injection or by intravenous infusion. An effective amount is from about 1 to 100 mg per kg per day. Parenteral solutions are prepared by dissolving ~e compounds used to practice the method claimed in this invention in water and filter sterilizing the solution before placing in a suitable sealable vial or arnpule. Parenteral suspensions are prepared in substantia}ly the same way except a sterile suspension vehicle is used and the compounds used to 10 practice the method claimed in this invention are sterilized wi~h ethylene oxide or suitable gas before it is suspended in the vehicle.
The exact route of administration, dose, or frequency of administration would be readily determined by those skill in the art and is dependant on the age, weight, general physical condition, or other clinical symptoms specific to the patient to be treated.
Patients to be treated would be those individuals: I) infected with one or more than one strain of a human immunodeficiency virus as determined by the presence of either measurable viral antibody or antigen in the serum and 2) having either a symptomatic AIDS defining infection such as i) disseminated histoplasmosis, ii) isoporiasis, iii) bronchial and pulmonary candidiasis including pneumocystis pneumonia, iv) non-Hodgkin's Iymphoma, or v) Kaposi's sarcoma and being less than sixty years old; or having an absolute CD4 Iymphocyte count of less than 200/mm3 in the peripheral blood. Treatment would consist of maintaining an inhibitory level of the compounds of this invention in the patient at all times and would continue until the occurrence of a second symptomatic AIDS defining infection indicates alternate therapy is needed.
Preparation 1: Preparation of N-poly-(o-hydroxybenzyl) Neomycin B
Part A:
12.33 g. ~20mM) of Neomycin B free base is dissolved in 25 rnl. of water. The solution is diluted with 50 ml of methanol. The resulting solution is mixed with an agueous methanol solution of salicylaldehyde (SAL; salicylic aldehyde; 2-hydroxybenzaldehyde) consisting of 17.1 g. (140 mM) of the SAL, 100 ml methanol and 200 ml water).
The resulting solution is heated to boiling, then allowed to stand at room temperature for 48 hours. The mixture is heated to boiling and 250 ml of hot water added and ~he mixture refrigerated at 0-5 C. for 24 hours. The supernatant is decanted and the resinous product dissolved in the methanol:dioxane (80:20), slurried with 2 g. of DARCO G60 charcoai and filtered though diatomaceous earth (Celite). The filtrate is added with stirring to 300 ml of ether to precipitate the desired Neomycin B Schiff base. The mixture is refrigerated overnight, and the supernatant decanted and the solid dried in vacuo.
WO 92/~2530 P(~/US9l/04849 3~ ~-Part B:
The product of Part A is dissolved in 200 ml methanol and hydrogenated at room temperaNre at 50 psi initial pressure for 24 hours with 1.0 g of PtO2 (Adarns' catalyst). The pressure drop is 7.0 psi per 100 mM. The product is fil~ered through diatomaceous earth (Celite) S and the filtrate concentrated 1n vacuo ~o about 50 ml and resulting concentra~e added slowly to 300 ml of ether. The resulting mixture is refrigerated overnight, fill~ered/decanted and the solid product dried initially at room temperan~re in vacuo for several days and finally overnight at 50 C. in a ~acuum oven to yield 9.5 g of N-poly-(o-hydroxybenzyl) Neomycin B. Analytical resul~s: N, 6.~8;
eq. wt. 216.1; Molecular Weight (MW) 1296.6; ~R 3260, 1605, 1586, 1485, 1045, 1030, 750.
Without further elaboration, those skilled in the art can practice the present invention to its fullest extent. The following detailed examples further describe how to use ~he compounds claimed in this invention to treat humans infected with one or more than one strain of a human immunodeficiency virus. These exdmpl~s are merely illustrative and are not limitations of the preceding disclosure. Those skilled in the art will promptly recognize appropriate Yariations from 15 the examples. In each exarnple, any compound claimed in this invention could replace the compound used in the particular example.
Example 1 Hard Gelatin Capsules One thousand two-piece hard gelatin capsules for oral use, each capsule containing 50 mg of N-poly-(o-hydroxybenzyl) Neomycin B, are prepared from the following:
N-poly-(o-hydroxybenzyl) ~leomycin B 50 grn Lactose 100 gm Cornstarch 20 gm Talc 20 gm Magnesium Stearate 2 gm The N-poly-(o-hydroxybenzyl) Neomycin B is added to the o~her ingredients, mixed and encapsulated in the usual manner.
Example 2 Tablets One thousand tablets, each containing 50 mg of N-poly-(o-hydroxybenzyl) Neomycin B are prepared f~om the following:
N-poly-(o-hydroxybenzyl) Neomycin B 50gm Lactose 75 gm Comstarch 50 gm Magnesium Stearate 4 gm I:ight liquid petrolatum S gm ;, ' ' ' ':
' wo92/û2530 2 0 8 5 3 6 7 Pcr/US9l/04849 The N-poly-(o-hydroxybenzyl) Neomycin B is added to the other ingredients, mi%ed and slugged. The slugs are broken down by forcing through a number sixteen screen. The resulting granules are then compressed into tablets.
Example 3 Parenteral solution A sterile aqueous solution for parenteral intravenous injection containing 150 mg of 1-(~
chlorobenzoyl)-N-poly-(o-hydroxybenzyl) Neomycin B in one liter of solution is prepared from the following:
N-poly-(o-hydroxyben~yl) Neomycin B 150 mg Water for injection, qs 1000 mg The N-poly-(o-hydroxybenzyl) Neomycin B is sterilized, added to the sterile water, filled into sterile containers and sealed.
The utility of this invention is demonstrated by the ability of N-poly-(o-hydroxyben7yl) Neomycin B to practice the method claimed in this invention to inhibit HIV-I tat media~ed transactivation, enzyme essentia~ for human immunodeficiency virus replication. HIV contains various genes, including ntatn which encodes a trans-activator (TAT,\ that functions in the infected cell by increasing the levels of steady-state viral mRNA as well as the transl$ional ~r~ilization of this mRNA. TAT ;s both essential for HIV replication and not structurally related to any norrnal cellular protein. nA Rapid, Quantitative Bioassay Based on the Human Immunodeficiency Virus Trans-Activatorn is described in AIDS RESEARCH AND HUMAN RETROVIRUSES, Vol. 5, Number 5, 1989, pp 507-516. The results (% Inhibition in sCD4 levels) in Table 1.
The anti-HlV utility of this invention is further demonstrated by determining the levels of HIV p24 and HIV RNA in culture supernatants of human peripheral blood Iymphocytes (PBLs) 3 and 4 days after HIV infection. The HIV infectivity experiments were conducted in primary cultures of (PBLs) with HTLV-llIB. Cultures were infected in triplicate with HIV, Compound I
Z5 added, and 3 to 4 days after infection the levels of HIV p24 synthesized and released were quantified by an enzyme-linked immunosorbent assay (ELISA) ~R. Maiolini and R. Masseyeff, 1.
Irnmunol. Methods 8, 223 (1975)] using a monoclonal capnlre antibody to HIV p24 (DuPont, Wilmington, DE) and recombinant HIV p24 calibration standard (MicroGenSys, West Haven, CI ).
The a nounts of HIV RNA synthesized in these infected cultures were also determined by hybridization analysis with a HlV-specific -32P labeled probe. The absolute HIV RNA levels were determined by norrnalizing hybridization values to values obtained from a standard preparation of HIV RNA hybridization .n pa.allel. The results are reported in Table 11.
The proliferation of mitogen-stimul3ted PBLs 4 days after exposure to Compound I was quantified relative to nondrug-treated controls to ensure that the infectivity experiments were conducted a~ non-cytotoxic concentrations of drug. At 40 ug/ml of Compound 1 did not inhibit PBL proliferation. The results are reported in Table In.
.
WO 92t02530 PCr/US91/04849 ~, ~
CHART I
5 Compound No. l N CH R
N0~ C~2 CN2 R~CH2NH _~ NH
,~
HO-CH2 o HO
R = 43 H
' WO 92/02530 PCI/IJ~;91/~4849 ' -7- 2V~367 TABLE I
INHIBITION OF HIY-1 tat MEDIATED TRANSACTIVATION
, ~;
uglml sCD4 in Culture Supernatant % Inhibition (ug/ml) in sCD4 Levels . 10 20 6.48 83 22. 15 43 .~ .
43.8 0 . .
0 38.7 o .
.
, "
.
. . ~
S i TABLE Il Inhibition of HIV-I Replication of U~466 % Inhibition U 6466 ~Mg/rnl) p24 HIV RNA
Day Day _ 4 86 8 l 82 66 0.4 22 0 12 9 0.04 20 4 23 17 ... .
.
:
WO 92/02530 PCr/US91/04849 9 2~3~7 U 6466 (ug/ml)% Cell Viability 0.4 g5 0.04 g6 .
~. :
Claims (10)
1. A method for inhibiting the replication of a human retrovirus (HRV) in a human infected with one or more than one strain of a human retrovirus comprising the systemic administration of an amount effective to inhibit the replication of the HRV of a N-poly-(o-hydroxybenzyl) Neomycin B of Formula I
where R is -(2-hydroxy)phenyl; or salts and/or hydrates thereof, to said human.
where R is -(2-hydroxy)phenyl; or salts and/or hydrates thereof, to said human.
2. A method according to Claim 1 where the human is infected with human immunodeficiency virus type I.
3. A method according to Claim 1 where the human is infected with human immunodeficiency virus type II.
4. A method according to Claim 1 where the administration is oral.
5. A method according to Claim 1 where the administration is parenteral.
6. A method according to Claim 1 where the compound has Molecular Weight (MW) of about 1296.6.
7. A method of treating a human having acquired immunodeficiency syndrome comprising the systemic administration of an effective inhibitory amount of a N-poly-(o-hydroxybenzyl) Neomycin B of Formula I
where R is -(2-hydroxy)phenyl; or salts and/or hydrates thereof, to said human.
where R is -(2-hydroxy)phenyl; or salts and/or hydrates thereof, to said human.
8. A N-poly-(o-hydroxybenzyl) Neomycin B of Formula I:
wherein R is -(2-hydroxy)phenyl; or salts and/or hydrates thereof.
wherein R is -(2-hydroxy)phenyl; or salts and/or hydrates thereof.
9. A compound according to Claim 8 having a molecular weight (MW) about 1296.6.
10. Use of a N-poly-(o-hydroxybenzyl) Neomycin B of Formula I
where R is -(2-hydroxy)phenyl, or salts and/or hydrates thereof, to prepare a medicament for inhibition of a human retrovirus (HRV) in a human infected with one or more strain of a human retrovirus.
where R is -(2-hydroxy)phenyl, or salts and/or hydrates thereof, to prepare a medicament for inhibition of a human retrovirus (HRV) in a human infected with one or more strain of a human retrovirus.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US55985190A | 1990-07-30 | 1990-07-30 | |
| US559,851 | 1990-07-30 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2085367A1 true CA2085367A1 (en) | 1992-01-31 |
Family
ID=24235298
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002085367A Abandoned CA2085367A1 (en) | 1990-07-30 | 1991-07-16 | Hiv-transactivators to treat aids |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP0541597A1 (en) |
| JP (1) | JPH05509306A (en) |
| AU (1) | AU640511B2 (en) |
| CA (1) | CA2085367A1 (en) |
| WO (1) | WO1992002530A1 (en) |
Families Citing this family (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5534408A (en) * | 1993-09-24 | 1996-07-09 | University Of Massachusetts Medical Center | 2-deoxystreptamine aminoglycoside inhibition of HIV RRE/Rev binding |
| JPH08502966A (en) * | 1992-10-23 | 1996-04-02 | ユニバーシティー・オブ・マサチューセッツ・メディカル・センター | Small molecule inhibition of RNA / ligand binding |
| PT1385540E (en) * | 2001-05-10 | 2011-10-21 | Wyeth Llc | Composition and method for increasing cell density in cell cultures infected with lentivirus |
| US20050143328A1 (en) * | 2003-10-31 | 2005-06-30 | Steele Philip M. | Composition and treatment for envelope virus infections |
| JP2008519061A (en) | 2004-11-05 | 2008-06-05 | アイシス ファーマシューティカルズ， インコーポレーテッド | Antibacterial 2-deoxystreptamine compound |
| WO2007028012A2 (en) * | 2005-09-01 | 2007-03-08 | Isis Pharmaceuticals, Inc. | Antibacterial 6'-n-modified 4,5-substituted aminoglycoside analogs |
| AU2006320374B2 (en) | 2005-12-02 | 2012-08-30 | Isis Pharmaceuticals, Inc. | Antibacterial 4,5-substituted aminoglycoside analogs having multiple substituents |
| WO2010030704A2 (en) | 2008-09-10 | 2010-03-18 | Achaogen, Inc. | Antibacterial aminoglycoside analogs |
| WO2010030690A1 (en) | 2008-09-10 | 2010-03-18 | Isis Pharmaceuticals, Inc. | Antibacterial 4,6-substituted 6', 6" and 1 modified aminoglycoside analogs |
| WO2010042850A1 (en) | 2008-10-09 | 2010-04-15 | Achaogen, Inc. | Antibacterial aminoglycoside analogs |
| WO2010042851A1 (en) | 2008-10-09 | 2010-04-15 | Achaogen, Inc. | Antibacterial aminoglycoside analogs |
| CA2777107A1 (en) | 2009-10-09 | 2011-04-14 | Achaogen, Inc. | Antibacterial aminoglycoside analogs |
| JP2013542992A (en) | 2010-11-17 | 2013-11-28 | アカオジェン インコーポレイテッド | Antibacterial aminoglycoside analogues |
-
1991
- 1991-07-16 JP JP3512462A patent/JPH05509306A/en active Pending
- 1991-07-16 WO PCT/US1991/004849 patent/WO1992002530A1/en not_active Application Discontinuation
- 1991-07-16 CA CA002085367A patent/CA2085367A1/en not_active Abandoned
- 1991-07-16 AU AU82300/91A patent/AU640511B2/en not_active Ceased
- 1991-07-16 EP EP91913197A patent/EP0541597A1/en not_active Ceased
Also Published As
| Publication number | Publication date |
|---|---|
| AU8230091A (en) | 1992-03-02 |
| EP0541597A1 (en) | 1993-05-19 |
| JPH05509306A (en) | 1993-12-22 |
| WO1992002530A1 (en) | 1992-02-20 |
| AU640511B2 (en) | 1993-08-26 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FZDE | Discontinued | ||
| FZDE | Discontinued |
Effective date: 19980716 |