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
  • A61K31/66—Phosphorus compounds
• • 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

Definitions

• the present invention relates to the use of at least one isomer inositoltrisphosphate (IP_) for the preparing of a medicament effective against retroviral diseases.
• IP_ inositoltrisphosphate
• HIV Acquired immunodeficiency syndrome
• HIV human immunodeficiency virus
• type 1 and type 2 HIV-1 and HIV-2
• HIV which is the aetiological agent for AIDS is a nononcogenic, cytopathic retrovirus of the lentiviru ⁇ subfamily.
• Retro ⁇ viruses have their genetic material in the form of RNA, ribonucleic acid, instead of DNA, deoxyribonucleic acid. In order to turn their RNA into DNA, retroviruses have a special enzyme called reverse transcriptase.
• the HIV-virus 100 nm in diameter is covered by an envelope, and contains a surface glycoprotein as well as an internal cylindrical core.
• the envelope is formed by phospholipids and glycoproteins and the core contains the genome and several enzymes.
• HIV infects human cells primarily by binding to CD4 receptors on the surface of susceptible cells. This binding is mediated by the oligomeric envelope glycoprotein (gp) of HIV and the receptor on the target cell surface followed by the fusion between the viral envelope and the plasma membrane.
• the post- binding events which lead to membrane fusion are poorly understood but presumably include a conformational change in the envelope protein which exposes the hydrophopic amino terminus of a gp 41 envelope protein in the fusion reaction.
• the binding of the HIV-1 envelope glycoprotein, gp 120, to the cellular receptor is the first step in HIV infection.
• CD4 independent mechanism of infection There are reports of HIV infection in a number of CD4-negative cells in vitro (Harouse, J.M. et al., J. Virol, 63, 2527, 1989, Zachar, V.B. et al. J. Virol, 65, 2102, 1991) .
• AIDS was first recognized in 1981 in young, homosexual men from the U.S. with opportunistic infections, Kaposi's sarcoma, and primary CNS lymphoma. Although unusual cases of systemic malignant lymphoma were also recognised at that time, statistically significant increases were not apparent until 1985. Victims also suffered from other opportunistic infections, caused by microorganisms that are ubiquitous but ordinarily not able to cause disease. Indeed, the infections and cancers seen in AIDS patients were previously known only in people born with certain defects in their immune system.
• HIV infection has been classified into three distinct stages: the acute phase, lasting weeks, the chronic phase, lasting years, and the final phase of crisis (generally referred to as AIDS) lasting months to years.
• AIDS is an unique disease. No other known infectious disease causes comparable harm by directly attacking the human immune system.
• HIV Once in the human body the virus attacks the cells that usually defend the body against infectious diseases. These cells include monocytes, macrophages and dendritic cells, so called antigen presenting cells (APC) . Furthermore HIV can remain hidden in cells latent for months or years. A third difficulty is that HIV is extraordinary variable in its genetic make-up.
• zidovudine was found to have in vitro activity against the human immunodeficiency virus, HIV.
• the in vitro and clinical activity of zidovudine is not disputed, but there is considerable debate when to initiate treatment (J.G. Bartlett,: New Engl. J. of Med., 329, 351, 1993; Cooper D.A. et al.: New Engl. J. of Med., 329, 297, 1993).
• AZT can have serious side effects such as anaemia, and in people using the drug, HIV frequently mutates to produce strains that are unaffected by it.
• nucleoside analogues are on trial but these will probably only be of limited usage as the mode of action including side effects reinforcee those of zidovudine.
• AZT The function of AZT is to block the action of the enzyme reverse transcriptase of HIV which stops the virus from replicating in the cells.
• Retroviruses vary at a notariously high rate resulting in rapid appearance of HIV-1 strains which are resistant against drugs and antibodies given to the patient.
• targets for therapeutic intervention are several other stages in the HIV replicate cycle.
• One such target is the HIV protease which is essential for the assenbly of fully infections HIV particles.
• -It should have high therapeutic indices -It should prevent the prolification of the HIV virus by preventing it from penetrating those cells in which the virus reproduces, or preventing it from reproducing inside these cells.
• the half-life of the drug should be long enough to allow and administration once or twice daily.
• the compound should be easily identified in the blood in order to establish individual dosage.
• the drug should effect the target cell in the same form as it is administered and not only as a result of metabolic activity.
• the drug should be teratologically safe.
• IP_ for the preparing of a medicament effective against retroviral diseases.
• the medicament is intended to be used for preventing, alleviating and combatting acquired immunodeficiency syndrome (AIDS) and AIDS-related diseases.
• AIDS acquired immunodeficiency syndrome
• the medicament can also be used against other conditions caused by retroviruses.
• the medicament exerts significant inhibitory effects against retroviral infections without side-effects which is very beneficial for the patient. Furthermore the medicament is also intended to be used against viral diseases caused by other enveloped viruses.
• the medicament is intended to be used against viral diseases caused by cytomegalovirus and different types of herpes virus.
• IP_ The production of IP_ and the isolation of the different isomers thereof are disclosed in the US patent No 4.777.134.
• the IP_ isomers can also be produced by synthetic methods, chemically or enzymatically, starting with e.g. inositol and a phosphorus source.
• microbiological production methods including hybrid DNA-techniques of IP are also suitable.
• IP_ and the different isomers thereof are disclosed in the US patent No 4.735.936 and the US patent No. 4.797.390.
• the medicament used according to the invention exists in unit dosage form.
• Tablets, granules or capsules are suitable administration forms for such unit dosage.
• tablets and granules can easily be surface treated such as to provide an enteric coating to prevent an uncontrolled hydrolysis in the stomach and to bring about a desired absorption in the intestine.
• Other suitable administration forms are slow release and trans- dermal administration.
• a usual pharmaceutically acceptable additive, excipient and/or carrier can be included in the medicament.
• the tablets or granules can also contain a disintegrant which causes the tablets or the granules, respectively, to disintegrate easily in the intestine.
• suspensions comprising the compound can be preferably used as administration form.
• the medicament can also consist as such of IP solely without any additive, excipient or carrier.
• the medicament can be free of other inositol phosphates, IP , ⁇ p 2 ' IP 4 ' IP 5 and IP 6 * Accor dingly, the mixture of IP isomers can have a purity of 90-100 %, such as 93-100 % or preferably 95-100 %.
• the medicament can consist of or comprise one or more specific IP isomers, each present in substantially pure form.
• the different isomers can be isolated from each other in substantially pure form, which means that they have a purity of 80-100 %, such as 82-100 % or 85-100 %, preferably 90-100 %. Since the isomers can be produced in pure form they can be mixed in any proportion, of course.
• the medicament can consist of IP , wherein said IP- is provided by at least one of IP fi/ IP or IP and a degradative substance such as an enzyme suitable to form IP-. It is in most cases suitable that the IP -isomer or isomers used for the preparing of the medicament according to the invention are present in salt form in order not to affect the mineral balance negatively.
• the salt should preferably consist of a sodium, potassium, calcium zinc or magnesium salt or a mixture of two or more of these salts.
• the medicament contains a surplus or an extra addition of at least one pharmaceutically acceptable salt of calcium, zinc or magnesium with a mineral acid or organic acid. This is especially valuable for elderly persons who are often deficient in these minerals.
• the preferred dosage for humans falls within the range of 0.1 to 1000 g, especially 0.1-200 mg IP_/day/kg body weight.
• the medicament usually contains 0.01-1.5 g, such as 0.05-1.3 g or preferably 0.1-1 g of IP 3 per unit dosage.
• composition used according to the present invention contains at least one, sometimes two or more of the following substances, which correspond to the essential IP_-isomer or isomer mentioned above: D-myo-inositol-l,2,6-trisphosphate of the formula
• X is hydrogen, at least one univalent, divalent or multivalent cation, or a mixture thereof, n is the number of ions, and z is the charge of the respective ion;
• L-myo-inositol-l,3,4-trisphosphate of the formula
• n ranges between 6 to 1 inclusive and z ranges from 1 to 6 inclusive.
• n is between 3 to 6 inclusive and z is 3, 2 or 1.
• D-myo-inositol-1,2,6-trisphosphate is preferred.
• inositol trisphosphate isomers that may be utilized in the present invention as the active IP- ingredient in the composition have the structural formula
• inositol trisphosphate compound is defined by the structural formula (I) where three of R., R , R 5 , R , R and R are hydroxyl and the remaining three are phosphate and R , R , R , R , R and R are hydrogen.
• inositol trisphosphates is defined by the structural formula (I) where three of Rl. , R_j, R c o l R protagonist/, R Q y and
• R 1? are hydroxyl and the remaining three are phosphate and ⁇ 2 , R 4 , R 5 , R 8 , 10 and R 1:L are hydrogen.
• Still another group of inositol trisphosphates is defined by the structural formula (I) where three of R 1. , R3_, RD R , Ro 0 , R.It and R. shall are hydroxyl and the remaining three are phosphate and R , R , R , R , R and R are hydrogen.
• inositol trisphosphates is defined by the structural formula (I) where three of R , R , R,., R , R_ and R are hydroxyl and the remaining three are phosphate and R , R , R , R , R and R are hydrogen.
• Still yet another group of inositol trisphosphates is defined by the structural formula (I) where three of R 1. , RJ_, R c o , Ro 0 ,
• R- and R are hydroxyl and the remaining three are phosphate and R , R , R , R , R and R . are hydrogen.
• inositol trisphosphates is defined by the structural formula (I) where three of R , R , R fi , R , R and R are hydroxyl and the remaining three are phosphate and R , R , R 5 , R , R g and R . are hydrogen.
• inositol trisphosphates is defined by the structural formula (I) where three of R , R , R , R , R. _ and R.. are hydroxyl and the remaining three are phosphate and R , R., R g , R ? , R and R are hydrogen.
• inositol trisphosphates is defined by the structural formula (I) where three of R , R , R , R , R g and R.. are hydroxyl and the remaining three are phosphate and R , R., R , R g , R and R are hydrogen.
• Particular inositol trisphosphate compounds within the contemplation of the above formula include compounds having the structural formula (I) where
• R Pain and R are phosphate, R , R and R. are hydroxyl and
• Rfest R , R 6 , R , R g and R.- are hydrogen
• R, R and R are phosphate, R , R and R_ are hydroxyl and R 4 , R 6 , R g , R 9 and R 2 are hydrogen;
• R and R are phosphate, R , R and R are hydroxyl and R 4 , R 6 , R 8 , R g and R 12 are hydrogen;
• R, R 5 and R are phosphate, R , R and R are hydroxyl and R, R , R-, R , R g and R 12 are hydrogen;
• R Nitrogen and R are phosphate, R , - and R- are hydroxyl and R , R , R g and R 12 are hydrogen;
• R and R are phosphate, R , R and R are hydroxyl and
• R, R., Rg, R , R g and R- 2 are hydrogen
• R- and R are phosphate, R , R and R 12 are hydroxyl and
• R, R 4 , R 5 , R 8 , R 10 and R ⁇ are hydrogen;
• R , R_ and R are phosphate, R , R and R are hydroxyl and R .
• R- , R Q , R-_ and R are hydrogen;
• R 5 and R g are phosphate, R , R and R are hydroxyl and R 4 , R g , R hinder, R g and R are hydrogen;
• R, R 3 and R 2 are phosphate, R , R and R are hydroxyl and R 4 , R g , R_, R g and R are hydrogen;
• R, R 3 and R 5 are phosphate, R g , R 1Q and R are hydroxyl and
• R, R 4 , R g , R_, R g and R are hydrogen
• R, R 5 and R g are phosphate, R 3 , R and R are hydroxyl and
• R, R 4 , R g ,R_, R and R - are hydrogen
• R 5 and R ⁇ 2 are phosphate, R 3 , R and R are hydroxyl and R 4 , R g , R_, and R are hydrogen;
• R 3 and R are phosphate, R , R and are hydroxyl and R 4 , R_, R_, R- 0 and R are hydrogen;
• R, R_ and R are phosphate, R , R and R are hydroxyl and R 4 , R 5 , R g , R and R. - are hydrogen;
• R and R are phosphate, R , R and R are hydroxyl and R 3 , R g , R restroom, R and R 11 are hydrogen;
• R. R 5 and R are phosphate, R , R and R are hydroxyl and
• Rr R 4 ,R , R , R and R ⁇ 2 are hydrogen
• R, R_ and R are phosphate, R , R and R are hydroxyl and R 4 , R g , R_, R g and R are hydrogen; R ⁇ R 3 and R 5 are phosphate, R ? , R g and R l are hydroxyl and
• R 2 R 4 , R 6 , Rg, R 10 and R 12 are hydrogen
• R l R 3 and R 12 are phosphate, R , R and R are hydroxyl and R 2 R 4 , R g , R ? , R 1Q and R 11 are hydrogen;
• R l R 3 and R g are phosphate, R 5 , R g and R are hydroxyl and R 2 R 4 , Rg, R_, R 10 and R. are hydrogen;
• R 3 R 5 and R ⁇ 2 are phosphate, R , R and R are hydroxyl and R 2 R 4 , R g , R_, R and R- - are hydrogen;
• R l R 5 and R g are phosphate, R 3 , R g and R are hydroxyl and R 2 R 4 , Rg, R restroom, R. and R are hydrogen;
• R l R 5 and R ⁇ 2 are phosphate, R 3 , R and R are hydroxyl and R 2 R 4 , R g , R_, R 10 and R. are hydrogen;
• R l R 3 and R g are phosphate, R 5 , R g and R are hydroxyl and R 2 R 4 , R g , R_, R 10 and R.. are hydrogen;
• R 3 R 5 and R are phosphate, R ⁇ R g and R are hydroxyl and R 2 R 4 , R g , R_, R 10 and R. are hydrogen;
• R 5 R g and R are phosphate, R , R and R are hydroxyl and R 2 R 4 , R g , R restroom, R 1Q and R are hydrogen;
• R l R g and R are phosphate, R 3 , R 5 and R are hydroxyl and R 2 R 4 , R g , R_, R._ and R are hydrogen;
• R l R g and R are phosphate, R , R and R are hydroxyl and R 2 R 4 , R g , R_, R 10 and R l are hydrogen; R, R g and R 12 are phosphate, R 1 , R 3 and R Q are hydroxyl and
• R, R 4 , R g , R ? , R 1Q and R ⁇ are hydrogen;
• R, R g and R are phosphate, R 3 , R and R are hydroxyl and R 4 , R g , R ? , R 1Q and 1 are hydrogen;
• Rg and R g are phosphate, R ⁇ R 3 and R are hydroxyl and R 4 , R g , R ? , R 10 and R ⁇ 1 are hydrogen;
• R 3 R 8 and R g are phosphate, R , R 5 and R are hydroxyl and R 2 R 4 , R g , R ? , R 10 and R ⁇ 1 are hydrogen;
• R, R g and R ⁇ 2 are phosphate, R 1 , R 5 and R g are hydroxyl and
• R, R 4 , R g , R 7 , R _ and R. are hydrogen;
• R g and R 12 are phosphate, R ⁇ , R and R are hydroxyl and R 4 , Rg, R ? , R _ and R are hydrogen;
• R g and R are phosphate, R., R and R 5 are hydroxyl and
• R. R 4 , R g , R ? , R - and R are hydrogen.
• Example l shows the manufacturing of a solution of IP for intravenous admini ⁇ stration and Example 2 and 3 demonstrate the effect of IP to inhibit infection induced by different HIV-cell lines.
• Example 4 illustrates the ability of IP to counteract HIV- infection induced by clinical isolates from HIV-patients.
• IP_ D-myo-inositol-1,2,6- trisphosphate
• IP 3 D-myo-inositol- 1,2,6-trisphosphate
• Quadriplicates of the suspensions were transferred to cell culture plates and were cultured for 4 days at 37°C.
• One measurement of the degree of infection is the expression of the antigen gp-120 on the cells.
• This HIV-antigen production was measured by an antibody-ELISA-technique and the results obtained for the different suspensions comprising IP were expressed relative to the suspension serving as a control.
• IP- D-myo-inositol-1,2,6-trisphosphate
• HIV-MN type code of HIV-virus
• Triplicates of the suspensions were transferred to cell culture plates and were cultured for 4 days at 37°C.
• the degree of infection was measured as the expression of the antigen gp-120 on the cells.
• the assay was performed using an antibody-ELISA-technique and the results obtained for the two suspensions comprising IP 3 were expressed relative to the suspension serving as a control.
• the data show a counteractive effect of IP against this type of HIV-induced infection.
• Viral isolates from HIV-infected patients were used in order to induce an infection by adding peripheral blood mononuclear cells (PBMC) to a medium containing different concentrations of the sodium salt of D-myo-inositol-1,2,6-trisphosphate (IP ) 25 CCID (50 % cell culture infections dose) of PBMCs from HIV-infected patients were incubated with three different concentrations of IP 3 ; 0.125 mg/ml, 0.250 mg/ml and 0.5 mg/ml. A fourth preparation, without any IP served as a control.
• the growth medium consisted of 10 % fetal calf serum, 2 ⁇ M glutamine, 100 IU/ml penicillin, 100 IU/ml streptomycin and 20 ⁇ g/ml gentamicin.
• PBMCs from healthy donors were then added to the prepara ⁇ tion. Before addition, these cells were stimulated for three days with phytohemagglutinin (PHA) .
• PHA phytohemagglutinin
• 0.5 x 10 PHA-stimulated PBMCs were added to each preparation, followed by incubation for 3 hours at 37°C. After extensive washing the cells were resuspended in growth medium supplemented with 10 IU/ml of interleukin-2 and seeded in quadruplicates of 100.000 cells in a 96-well microtite plate before further cultivation for 7 days.
• the HIV-antigen production was assayed at the seventh day using an ELISA-technique.
• the obtained values were norma ⁇ lized and are summarized in the following table:
• IP shows a strong effect to counteract the infection. At the concentration of 0.5 mg/ml of IP the infection is totally abolished.

Landscapes

• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Veterinary Medicine (AREA)
• Chemical & Material Sciences (AREA)
• Medicinal Chemistry (AREA)
• Pharmacology & Pharmacy (AREA)
• Animal Behavior & Ethology (AREA)
• General Health & Medical Sciences (AREA)
• Public Health (AREA)
• Epidemiology (AREA)
• Virology (AREA)
• Communicable Diseases (AREA)
• Oncology (AREA)
• Chemical Kinetics & Catalysis (AREA)
• General Chemical & Material Sciences (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Organic Chemistry (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Applications Claiming Priority (3)

Publications (1)

Family

ID=20391349

Family Applications (1)

Country Status (5)

Cited By (1)

Families Citing this family (1)

Family Cites Families (1)

• 1993
  • 1993-10-08 SE SE9303289A patent/SE501793C2/sv not_active IP Right Cessation
• 1994
  • 1994-10-06 JP JP7511669A patent/JPH09503519A/ja active Pending
  • 1994-10-06 AU AU78668/94A patent/AU7866894A/en not_active Abandoned
  • 1994-10-06 WO PCT/SE1994/000928 patent/WO1995010285A1/en not_active Application Discontinuation
  • 1994-10-06 EP EP94929713A patent/EP0724445A1/de not_active Withdrawn

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events