CA1339147C - Salt-type compounds, pharmaceutical compositions containing them and process for preparing same - Google Patents

Salt-type compounds, pharmaceutical compositions containing them and process for preparing same

Info

Publication number
CA1339147C
CA1339147C CA000593269A CA593269A CA1339147C CA 1339147 C CA1339147 C CA 1339147C CA 000593269 A CA000593269 A CA 000593269A CA 593269 A CA593269 A CA 593269A CA 1339147 C CA1339147 C CA 1339147C
Authority
CA
Canada
Prior art keywords
acid
salt
lysine
general formula
antiviral
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.)
Expired - Fee Related
Application number
CA000593269A
Other languages
French (fr)
Inventor
Peter Literati Nagy
Maria Boros
Istvan Racz
Jeno Szilbereky
Gyongyver Soos
Gabor Nemeth
Alan Pinter
Miklos Koller
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
BIOREX KFT
Original Assignee
BIOREX KFT
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by BIOREX KFT filed Critical BIOREX KFT
Application granted granted Critical
Publication of CA1339147C publication Critical patent/CA1339147C/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/64—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with substituted hydrocarbon radicals attached to ring carbon atoms, e.g. histidine
    • 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
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C227/14—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton from compounds containing already amino and carboxyl groups or derivatives thereof
    • C07C227/18—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton from compounds containing already amino and carboxyl groups or derivatives thereof by reactions involving amino or carboxyl groups, e.g. hydrolysis of esters or amides, by formation of halides, salts or esters
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C229/06—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton
    • C07C229/08—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to hydrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C229/22—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated the carbon skeleton being further substituted by oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C229/24—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having more than one carboxyl group bound to the carbon skeleton, e.g. aspartic acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C229/26—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having more than one amino group bound to the carbon skeleton, e.g. lysine
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/34—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton containing six-membered aromatic rings
    • C07C229/36—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton containing six-membered aromatic rings with at least one amino group and one carboxyl group bound to the same carbon atom of the carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C237/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups
    • C07C237/02—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton
    • C07C237/04—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C237/06—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being acyclic and saturated having the nitrogen atoms of the carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C279/00—Derivatives of guanidine, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups
    • C07C279/04—Derivatives of guanidine, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of guanidine groups bound to acyclic carbon atoms of a carbon skeleton
    • C07C279/14—Derivatives of guanidine, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of guanidine groups bound to acyclic carbon atoms of a carbon skeleton being further substituted by carboxyl groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C57/00—Unsaturated compounds having carboxyl groups bound to acyclic carbon atoms
    • C07C57/02—Unsaturated compounds having carboxyl groups bound to acyclic carbon atoms with only carbon-to-carbon double bonds as unsaturation
    • C07C57/03—Monocarboxylic acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D207/10—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D207/16—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Virology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Oncology (AREA)
  • Communicable Diseases (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • AIDS & HIV (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Epidemiology (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicinal Preparation (AREA)

Abstract

The invention relates to novel antiviral and immunostimulant salts of the general formula:

R-COO-AH+ (I) wherein R means a C18-24 alkyl group containing at least two double bonds; and "A" stands for an amino acid occurring in living organisms and/or a derivative thereof containing a carboxyl group substituted by a C1-4 alkyl group or an amino group or by an alkali metal cation. The invention further relates to a process for the preparation of the above salts.

Description

1~91~7 This invention relates to novel antiviral and immune system-stimulating salts and pharmaceutical compositions containing these salts, as well as a process for the preparation of the new salts.
The advantageous antiviral action of ~-3 poly-unsaturated fatty acids [5,8,11,14,17-eicosapentaenoic acid (hereinafter: EPA) and 4,7,10,13,16,19-docoahexaenoic acid (hereinafter: DHA)] has generated considerable attention.
Szads [Antimicrobial Agents and Chemoterapy 12, 523 (1977)]
showed by ln vitro experiments that polyunsaturated fatty acids, e.g. both EPA and DHA, are capable of inhibiting virus replication. The same fact was supported by Reinhardt et al. [J. of Virology 25, 479 (1978)] investigating the inhibition of the replication on PR 4 bacteriophage.
The antiviral effect of polyunsaturated fatty acids, including that of EPA and DHA, is described in detail in United States Patent No. 4,513,008. The effect of EPA
and DHA was compared in animal experiments on mice and guinea pigs to Acyclovir [9-(2-hydroxyethoxymethyl)guanine]
being the most frequently used antiviral agent at present.
It has been stated that compositions containing particularly DHA showed a more preferable action against herpes virus than Acyclovir.
A number of articles have been devoted in the literature to the antiviral effect of DHA and EPA, such as:
'Whitaker et al. [Proc. Natl. Acad. Sci. USA 76, 5919 (1979)], as well as Goodnight et al. [Arterioschlerosis, 2, :~339147 87 (1982)] and Yoshiaki [Biochimica et Biophysica Acta 793, 80 (1984)].
Prickett et al. [Immunology 46, 819 (1982)] showed that the humoral immune response was stimulated by eicosa-pentaenoic acid as an arachidonic acid analogue. Under anEPA-rich diet, the specific IgG and IgE production as a response give to egg albumin was increased in inbred Sprague-Dawley rats to a 4 to 8-fold value in comparison to the control. According to this paper, an enhanced antibody response was induced by the EPA-rich diet. Further on, there was shown by the investigations that EPA acts through an inhibition of the suppressive prostaglandin system whereby it is capable of inhibiting or correcting, respectively, the immune deficiency accompanying ageing and other pathological processes (autoimmune processes, tumourogenesis). These statements were supported by the articles of Kelley et al. [J. of Immunol. 134, 1914 (1985)]
as well as Homey et al. [Clin. Exp. Immunol. 65, 473 (1986)].
It has long been known on the basis of the in vivo investigations of Pearson et al. [Proc. Soc. Exp. Biol. Med.
79, 409 (1952)], that L-lysine has an effect in inhibiting the encephalomyelitis virus. Subsequently, Tankersley [J.
8act. 87, 609 (1964)] called attention to the fact that the replication of the herpes simplex virus (hereinafter: HSV) is inhibited by lysine in human cells under in vitro conditions. Based on human examinations, Kagan published ~ I
.~.

[The Lancet 1, 137 (1974)] that both oral and genital HSV-induced lesions very rapidly disappeared on treatment with L-lysine.
Griffith et al. [Dermatologica 156, 257 (1978)]
studied the therapeutic action of L-lysine in various doses on 45 patients infected by HSV I and HSV II, with varied treatment periodsn The age of the patients (predominantly women) varied between 8 and 60 years. It was noted that L-lysine exerted only a suppressive but not a healing effect on HSV.
The present invention is directed to novel, therapeutically effective salts and the use of these salts for preparing pharmaceutical compositions, which combine the advantageous therapeutical effects of the ~-3 unsaturated fatty acids with those of amino acids, such as in particular lysine, ornithine and histidine, and which show a stronger action than any antiviral agent thus far known.
The invention is based on the recognition that the salts obtained by the salt-formation of ~-3 unsaturated fatty acids, particularly EPA and DHA, with amino acids or their derivatives, respectively, particularly with lysine, possess a strong antiviral and immunostimulating effect.
Thus, the present invention relates to novel antiviral and immunostimulating salts of the general formula:

R-COO AH (I) C' - ~ 13~9147 wherein R means a C1824 alkyl group containing at least two double bonds; and "A" stands for an L-amino acid occurring in living organisms selected from L-tyrosine, L-lysine, L-histidine, L(+)-ornithine, L-alanine, L-proline, L-leucine, L-threonine, L-aspartic acid, L-arginine and L-serine.
According to the invention, these novel salts are prepared by reacting as base component an amino acid "A" as defined for the general formula (I), wherein the substituents are the same as defined above, with an acid of the general formu]a:

R-COOH (II) wherein R is the same as defined above, in a polar solvent.
The compounds of general formula (I) are preferably salts formed from ~-3 unsaturated fatty acids containing at least two double bonds with basic amino acids or their derivatives, respectively.
The eventual toxicity against a cell culture of the salt compounds according to the invention was studied with a salt formed from an ~-3 polyunsaturated fatty acid mixture (containing 27.6% of EPA and 44.6% of DHA) with L-lysine on the basis of its effect exerted on the propagationand morphology of Hep 2 cells (human epithelial tumour cell line). These experiments were carried out on a plastic I

13391~7 tissue-cultivating sheet containing 6x4 hollow spaces (holes with a bottom surface of 1.9 cm2 each).
A stock solution containing the test compound in a concentration of 10 mg/ml was prepared in Eagle MEM medium (manufactured by Serva GmbH Co., Heidelberg, German Federal Republic). An aliquot of the stock solution was diluted 10-fold, the solution obtained was diluted 2-fold and then, by repeating twice the 2-fold dilution of the latter solution, solutions with successively decreasing concentrations of the test compound (solutions Nos. 1 to 5) were prepared. The cells were treated with 1 ml each of a solution containing the active ingredient in the following concentrations:

Solution Concentration of the No. active ingredient ~g/ml 10, 000 2 1,000 The treatment was carried out for 1 hour, then the cultures were washed twice with buffered sodium chloride solution (phosphate-buffered saline, hereinafter: PBS), then a nutritive medium was added to the cultures. After 24 :hours and fixation by methanol, the cultures were dyed by ethanolic Giemsa solution (manufactured by Reanal, Budapest, Hungary) and the morphology of the cells was evaluated under ~ ,,.~

13311~7 a light microscope. It was noted that the test compound proved to be toxic only above a concentration of 1000 ~g/ml.
The virus replication-inhibiting effect was ~ ;ned on Hep 2 cells as described above. Type I of HSV
was used for infection. The concentration of the virus amounted to about 1000 PFU (plaque forming unit). Solutions containing the test compound in a concentration of 1000, 500 and 250 ~g/ml, respectively, were used and for the treatment 0.1 ml of solution was added to the undiluted virus suspension. The mixture was incubated at 37~C for 1 hour.
Then, the cells were treated by the virus pre-incubated with the test compound and the development of cytopathologic (hereinafter: CP) alterations was observed for 7 days.
It was noted that virus replication was directly inhibited by a 500 ~g/ml or 250 ~g/ml dose of the test compound, and the infective titre value of the cytopathologic dose (neg. log. CPD50) was 1.75, calculated for 0.1 ml, in comparison to the neg. log. CPD50 value of 4.5 of the untreated control virus culture. The value of the virus inhibition exceeds by two orders of magnitude that of the control. Under the same conditions, EPA and DHA did not show any significant virus inhibition; and lysine itself inhibited the virus replication to an extent of only about one order of magnitude.
Vaccine virus was also treated in an in vitro experiment carried out in the above manner. The infective titre value (neg. log. CPD50) of the test compound measured .~

13~9147 on vaccine virus proved to be 1.75 in comparison to the 5.5 value of the untreated control virus, i.e. the test compound showed a virus-inhibiting effect which was three orders of magnitude stronger than that of the control.
The in vitro action of the compounds according to the invention on the immune system was studied with respect to the activation of lymphocytes by polyclonal mitogens.
The blastic transformation of the lymphocytes was investigated in such a way that a lymphocyte cell population obtained on a Fico Uromiro gradient [Scand. J. Clin. Lab.
Invest. 21, 97 (1968)] was pipetted into the holes of flat-ground sheets, 25 ~g/ml of Concanavaline A (hereinafter:
Con A) (manufactured by Pharmacia, Uppsala, Sweden) and then a solution containing the novel compound according to the invention in a concentration of 0.1, 1.0 or 10 ~g/ml, respectively, were added to each of the parallel cultures.
A culture cont~in;ng 25 ~g/ml of Con A without the test compound was used as control. The sheets were maintained under an air atmosphere containing 5% of carbon dioxide at 37~C for 72 hours, and 0.4 ~Ci of H-thymidine was added to each sample after 64 hours, before ending the cultivation.
After 72 hours, the cultures were filtered through a glass filter, the filters were put into a scintillation cuvet and the radioactivity of each was measured in 5 ml of toluene solution by using a beta-counter device. The results are summarized in the following Table.

~3~9147 o . ~,, ~ ~j .. ..

~ ~ ~ ~ ~ U. ~ U
v .. .. ... .. . ..
.C U ~ UJ
U ,y ~ U Ul ; U
r :
o a) -I o o ~ o ~ ~
n ~ UJ

$ d~ In ~ w ~ ~ O
~" d' O ~ ~ ~ ~ ~~ o a~ ~ ~ ~ ~~ ~ ~D ~ ~ ~1 ~ ~ +l +l +l +l +l +l +l +l +l +l +l +l V ~ ~ t~ ~ ~w r~
~ ~ ~ o w ~ o~ ~ ~ ~ o o ~w ~ l ~w ~ ~ ~l ~l r~
C
. r~ ~
,.~ o ,,~ o ri~ o ~ ~ ~ ~ o ~-rl ~ o r.~ o ~i o ~ ~ o r.~ o r.
~ rC
V J~
O q~
C~ O ~
~r~l r.~ ~I
~ r~ ~ ~
r,~ r~
r~ ,x ~,~ ~ r~ r~
O O O
a ~ v ~
o -- ~ o-,~ r,~-- o o r.--l 'L ~ 1 ~ J
O :~ '' r-l r,-l ~ ~ ') O
- ~ UJ
-- ~ '~ Ul ~
UJ ~ :
Z 4 ~

J 13~9l~7 It is obvious from these P~m;nations that the compounds according to the invention and particularly the salt of lysine or tyrosine formed with a mixture of polyunsaturated fatty acids show significant test results, i.e. a strong immunostimulating effect, whilst the separate salt-forming components in themselves proved to have no or only a weak biological action.
~ s starting material comprising C1824 ~-3 unsaturated fatty acids as one component of the salts, it is suitable to use first of all oils which can be obtained from fish of the northern seas, such as salmon, codfish or sardine, or from their livers. However, oils arising from fresh-water fish can also be utilized. The ~-3 poly-unsaturated fatty acids are obtained from the above oils by using a known method [J. Am. Chem. Soc. 59, 117 (1982)].
The active compounds of general formula (I) can be transformed to capsules, tablets, dragées, suppositories or other pharmaceutical compositions formulated in a manner known per se by using carriers and/or additives, such as lactose, starch, magnesium stearate and the like, commonly used in the pharmaceutical industry.
For inhibiting the oxidation of the composition, the use of ~-tocopherol (vitamin E), glutathione or a traditional antioxidant, such as butylhydroxytoluene, is suitable.

1~91~7 The main advantages of the compounds according to the invention and of the pharmaceutical compositions prepared therefrom can be summarized as follows:
1. They can be used against acute virus infections; they are particularly preferable in the herpes virus infection for suppressing the infection at an early pointA
2. Owing to their immunostimulating effect, they can preferably be used against retroviruses, especially against the immunodeficiency syndromes (e.g. AIDS) induced by HTLV III and HTLV IV type viruses.
3. They contain exclusively native active agents which are essential from a biological viewpoint. Thus, they are useful for a prophylactic, long-lasting, cure-like treatment in case of danger of virus infection as well as in diseases of the immune system.
4. They also act internally. Thus, the inconvenient external treatment commonly used in antiviral therapy can be avoided.
The compounds and compositions according to the invention and their process of preparation are illustrated in detail by the following non-limiting Examples.

Example 1 To 164 g (1 mole) of L-lysine monohydrate dissolved in 500 ml of water at room temperature, 320 g (about 1 mole) of an ~-3 polyunsaturated fatty acid mixture ~ I

13391i7 (containing 27.6% of BPA, 44.6% of DHA and 0.1% of vitamin E) are added dropwise. The mixture is stirred under mild heating (at 40~C) and under nitrogen for 3 hours and then evaporated under reduced pressure (4 to 5.3 kPa) to obtain 465 g of a crystalline salt, m. p.: 188-195~C (with decomposition), the unsaturated fatty acid composition of which is identical to that of the starting mixture.

Example 2 155 g (1 mole) of L-histidine are dissolved in 450 ml of water at room temperature and 320 g (about 1 mole) of an ~-3 polyunsaturated fatty acid mixture (containing 27.6%
of EPA and 44.6% of DHA) are added dropwise to the solution over 5 minutes. The process of Example 1 is then followed to give 471 g of a crystalline substance, m. p.: 192-200~C
(with decomposition).

Example 3 The process described in Example 2 is followed, except that 133 g of L(+)-ornithine are used instead of L-histidine to obtain 449 g of a crystalline product, m. p.:
189-195~C (with decomposition).

Example 4 After dissolving 1.64 g (0.01 mole) of L-lysine monohydrate in 5 ml of water at room temperature, 3.02 g (0.01 mole) of eicosapentaenoic acid [manufactured by Sigma f~., . .~

1~39147 Co., St. Louis, USA under catalogue number E-7006 (1987)]
are added to the solution in small portions. The mixture is maintained at 40~C under nitrogen for 3 hours, then evaporated under a pressure of 4-5.3 kPa to give 4.9 g of a yellowish brown crystalline salt, m. p.: 194-196~C (with decomposition).

Example 5 The process described in Example 4 is followed, except that 3.28 g (0.01 mole) of docosahexaenoic acid [manufactured by Sigma Co., St. Louis, USA under the catalogue number D-6508 (1987)] are used instead of eicosapentaenoic acid, to give 4.85 g of a crystalline salt, m. p.: 195-198~C (with decomposition).
Example 6 The process described in Example 4 is followed, except that 1.55 g (0.01 mole) of L-histidine are used instead of L-lysine monohydrate to obtain 4.8 g of a crystalline salt, m. p.: 196-200~C (with decomposition).

Example 7 The process described in Example 4 is followed, except that 1.55 g (0.01 mole) of L-histidine instead of L-lysine monohydrate and 3.28 g (0.01 mole) of DHA instead of EPA are employed to give 3.78 g of a crystalline salt, m.
p.: 196-199~C (with decomposition).

....
, --Example 8 The process described in Example 4 is followed, except that 1.32 g (0.01 mole) of L(+)-ornithine are used instead of L-lysine monohydrate to give 4.6 g of a crystalline salt, m. p.: 190-193~C (with decomposition).

Example 9 The process described in Example 4 is followed except that 1.32 g (0.01 mole) of L(+)-ornithine instead of L-lysine monohydrate and 3.28 g (0.01 mole) of DHA instead of EPA are used to give 4.55 g of a crystalline salt, m. p.:
191-195~C (with decomposition).

Example 10 2 g (22 mmoles) of L-alanine are dissolved in a solution of 0.88 g (22 mmoles) of sodium hydroxide in 20 ml of water while stirring at room temperature. Then, 7.5 g (22 mmoles) of an ~-3 polyunsaturated fatty acid mixture (containing 27.6% of EPA, 44.6% of DHA and 0.1% of vitamin E) are added dropwise at 40~C to the above solution. The mixture is stirred under nitrogen at 40~C for 2 hours, then the solvent is distilled off under a reduced pressure of 4 to 5.3 kPa to give 10.4 g of a pale brown paste-like solid product, m. p.: 210-220~C.
In the following Examples 11 to 14, the process described in Example 10 is followed, except that the stated amino acid is used instead of L-alanine.

~ I

~, 13391~7 Example 11 L-Proline 1.5 g (13.0 mmoles) Water 40.0 ml Sodium hydroxide 0.52 g (13.0 mmoles) ~-3 fatty acid mixture 4.35 g (13.0 mmoles) (according to Example 10) A brown oily product is obtained in a yield of 6.3 g.

Example 12 L-Leucine 1.0 g (7.6 mmoles) Water 5.0 ml Sodium hydroxide 0.3 g (7.6 mmoles) ~-3 fatty acid 2.54 g (7.6 mmoles) A brown crystalline product is obtained in a yield of 3.70 g which liquifies in the air.

Example 13 L-Threonine 0.5 g (4.2 mmoles) Water 10.0 ml Sodium hydroxide 0.16 g (4.2 mmoles) ~-3 fatty acid 1.4 g (4.2 mmoles) A yellow crystalline product is obtained in a yield of 1.95 g, m. p.: 204-213~C (with decomposition).

~, I

1~39147 Example 14 L-Aspartic acid 1.0 g (7.5 mmoles) Water 40.0 ml Sodium hydroxide 0.6 g (15.0 mmoles) ~-3 fatty acid 2.5 g (7.5 mmoles) A yellow crystalline salt is obtained in a yield of 3.97 g, m. p.: 200~C (with decomposition).

Example 15 7.1 mmoles of sodium metal are dissolved in 20 ml of anhydrous ethanol, and the solution is cooled to between 0~C and 10~C and 1.5 g (7.1 mmoles) of L-arginine hydro-chloride are added. After stirring at room temperature for 20 minutes, the mixture is filtered and the filtrate is evaporated under reduced pressure. The evaporation residue is dissolved in a solution of 0.28 g (7.1 mmoles) of sodium hydroxide in 15 ml of water, then 2.37 g (7.1 mmoles) of an ~-3 polyunsaturated fatty acid mixture (containing 27.6% of EPA, 44.6% of DHA and 0.1% of vitamin E) are added. The mixture is then stirred at 40~C under nitrogen for 2 hours.
Thereafter, the solvent is distilled off under a reduced pressure of 4-5.3 kPa to give 4.05 g of a yellow crystalline salt, m. p.: 207-211~C (with decomposition).

Bxample 16 2.0 g (11.0 mmoles) of L-tyrosine are dissolved in a solution cont~;n;ng 0.44 g (11.0 mmoles) of sodium ~, 1339147 hydroxide in 20 ml of water and 20 ml of methanol at room temperature, then 3.68 g (11.0 mmoles) of an ~-3 poly-unsaturated fatty acid mixture (with the same composition as described in Example 1) are added dropwise to the above solution at 40~C. After stirring the reaction mixture under nitrogen at 40~C for 2 hours, the solvents are evaporated under a reduced pressure of 4 to 5.3 kPa to give a yellowish, solid powder-like crystalline salt residue in a yield of 6.18 g, m. p.: 150~C (with partial decomposition).
In the following Examples 17 and 18 the process described in Example 16 is followed, except that the salt is formed by using ~he indicated amino acid instead of L-tyrosine.

Example 17 L-Serine 1.0 g (9.5 mmoles) Methanol 10.00 ml Water 15.0 ml Sodium hydroxide 0.38 g (9.5 mmoles) ~-3 fatty acid mixture 3.17 g (9.5 mmoles) (according to Example 1) After evaporation, a pale brown crystalline product is obtained in a yield of 4.5 g, which decomposes over 175~C.

~, I

.1 13391i7 Example 18 Glutamine 2.0 g (13.77 mmoles) Methanol 20.0 ml Water 105.0 ml Sodium hydroxide 0.55 g (13.77 mmoles) ~-3 fatty acid mixture 4.60 g (13.77 mmoles) A brownish powder-like crystalline salt is obtained in a yield of 7.11 g, m. p.: 181-190~C.
Example 19 Preparation of a pharmaceutical composition in capsule form A salt mixture prepared as described in Example 1 is filled into hard gelatine capsules, capable of receiving 500 mg of active ingredient, using a known encapsulation process.

Example 20 Preparation of tal~lets Tablets are prepared from the salt mixture obtained as described in Example 1, each of which tablets contains the following components:

Salt mixture according to Example 1 500 mg Lactose 120 mg 25 Starch 63 mg Polyvinylpyrrolidone 3.5 mg Magnesium stearate 3.5 mg If desired, the tablets may be covered with a sugar coating by using a panning machine.

~,

Claims (12)

1. A novel antiviral and immunostimulating salt of the general formula:

R-COO-AH+ (I) wherein R means a C18-24 alkyl group containing at least two double bonds; and "A" stands for an L-amino acid occurring in living organisms selected from L-tyrosine, L-lysine, L-histidine, L(+)-ornithine, L-alanine, L-proline, L-leucine, L-threonine, L-aspartic acid, L-arginine and L-serine.
2. A salt as claimed in claim 1, wherein A
represents L-lysine or L-tyrosine.
3. A salt as claimed in claim 1, which comprises eicosapentaenoic acid (EPA) as acid component and L-lysine as base component.
4. A salt as claimed in claim 1, which comprises docosahexaenoic acid (DHA) as acid component and L-lysine as base component.
5. A salt as claimed in claim 1, which comprises a mixture of docosahexaenoic acid and eicosapentaenoic acid as acid component and L-lysine as base component.
6. An antiviral and immunostimulant pharmaceutical composition, which comprises a salt of the general formula (I) as defined in claim 1, in admixture with a pharmaceutically acceptable carrier or additive.
7. A process for the preparation of a novel salt of the general formula:

R-COO-AH+ (I) wherein R and A are as defined in claim 1, which comprises reacting as base component an L-amino acid occurring in living organisms and selected from L-tyrosine, L-lysine, L-histidine, L(+)-ornithine, L-alanine, L-proline, L-leucine, L-threonine, L-aspartic acid, L-arginine and L-serine with an acid of the general formula:

R-COOH (II) wherein R is as defined in claim 1, in a polar solvent.
8. A process as claimed in claim 7, wherein the acid of formula (II) is eicosapentaenoic acid and the amino acid is L-lysine.
9. A process as claimed in claim 7, wherein the acid of formula (II) is docosahexaenoic acid and the amino acid is L-lysine.
10. A process as claimed in claim 7, wherein the acid of formula (II) is a mixture of docosahexaenoic acid and eicosapentaenoic acid and the amino acid is L-lysine.
11. A process for preparing a pharmaceutical composition having antiviral and immunostimulant effect, which comprises admixing a salt of the general formula (I) prepared by the process as claimed in claim 7, with a pharmaceutically acceptable carrier and/or auxiliary agent, and transforming the thus-obtained mixture in the form of tablets, dragées, capsules or suppositories to a pharmaceutical preparation.
12. Use of the salts of the general formula (I) as claimed in claim 1 for preparing pharmaceutical compositions having antiviral and immunostimulant activity.
CA000593269A 1988-03-09 1989-03-09 Salt-type compounds, pharmaceutical compositions containing them and process for preparing same Expired - Fee Related CA1339147C (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
HU881131A HU199775B (en) 1988-03-09 1988-03-09 Process for production of formed by fatty acids salts of amin acids and medical compositions containing them
HU1131/88 1988-03-09

Publications (1)

Publication Number Publication Date
CA1339147C true CA1339147C (en) 1997-07-29

Family

ID=10952971

Family Applications (1)

Application Number Title Priority Date Filing Date
CA000593269A Expired - Fee Related CA1339147C (en) 1988-03-09 1989-03-09 Salt-type compounds, pharmaceutical compositions containing them and process for preparing same

Country Status (16)

Country Link
JP (1) JP2751068B2 (en)
KR (1) KR0140993B1 (en)
AT (1) AT398073B (en)
BE (1) BE1003663A3 (en)
CA (1) CA1339147C (en)
CH (1) CH678851A5 (en)
DE (1) DE3907688C2 (en)
ES (1) ES2010439A6 (en)
FI (1) FI93949C (en)
FR (1) FR2628419B1 (en)
GB (1) GB2216522B (en)
HU (1) HU199775B (en)
IT (1) IT1229563B (en)
LU (1) LU87470A1 (en)
NL (1) NL8900573A (en)
SE (1) SE508603C2 (en)

Families Citing this family (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH04279523A (en) * 1991-01-11 1992-10-05 Nisshin Flour Milling Co Ltd Fatty oil processed product for stimulating immunity
DE69231592T2 (en) * 1991-06-24 2001-06-21 Women's And Children's Hospital, North Adelaide METHOD AND COMPOSITIONS FOR TREATING MALARIA AND OTHER DISEASES
WO1995009622A1 (en) * 1993-10-06 1995-04-13 Peptide Technology Limited Polyunsaturated fatty acids and uses thereof
NL9401743A (en) * 1994-10-20 1996-06-03 Prospa Bv Salts of amino alcohols and pharmaceutical formulations containing them.
AUPM906594A0 (en) * 1994-10-26 1994-11-17 Peptide Technology Limited Synthetic polyunsaturated fatty acid analogues
US5639858A (en) * 1995-03-22 1997-06-17 Tularik, Inc. Human signal transducer and binding assays
HU227588B1 (en) * 2004-12-03 2011-09-28 Sinnex Mueszaki Fejlesztoe Es Tanacsado Kft Antiviral and immunostimulant pharmaceutical composition containing polyunsaturated fatty acid esters
WO2007079394A2 (en) * 2005-12-30 2007-07-12 Revance Therapeutics, Inc. Arginine heteromers for topical administration
PL3236782T3 (en) * 2014-12-23 2020-06-29 Evonik Operations Gmbh Process for increasing the stability of a composition comprising polyunsaturated omega-3 fatty acids
EP3248467A1 (en) * 2016-05-25 2017-11-29 Evonik Technochemie GmbH Method for preparing a composition containing omega-3-fatty acid-l-lysin-salts
EP3668498A1 (en) * 2017-08-15 2020-06-24 Evonik Operations GmbH Tablets with high active ingredient content of omega-3 fatty acid amino acid salts
US20230065371A1 (en) * 2020-01-30 2023-03-02 Silicycle Inc. Process for manufacturing solid neutral amino acid salts of polyunsaturated fatty acids

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS4887011A (en) * 1972-02-04 1973-11-16
US4513008A (en) * 1982-07-30 1985-04-23 The Vinoxen Company, Inc. Virucidal compositions and therapy
GB8327911D0 (en) * 1983-10-19 1983-11-23 Ciba Geigy Ag Salts as corrosion inhibitors
US4537772A (en) * 1984-05-02 1985-08-27 Merck & Co., Inc. Enhancing absorption of drugs from gastrointestinal tract using acylcarnitines
FR2587900B1 (en) * 1985-10-01 1988-10-07 Morelle Jean BASIC FATTY-AMINO ACID ASSOCIATIONS WITH EMOLLIENT, EMULSIFIING AND ANTIOXIDANT PROPERTIES FOR COSMETICS, DERMATOLOGY AND FOOD
JPS63230632A (en) * 1987-03-20 1988-09-27 Nippon Oil & Fats Co Ltd Agent for stimulating lipoxygenase metabolism
DE3726299A1 (en) * 1987-06-26 1989-02-23 Dietl Hans Fat emulsion for intravenous use

Also Published As

Publication number Publication date
DE3907688C2 (en) 1996-09-05
IT8919705A0 (en) 1989-03-09
FI891144A (en) 1989-09-10
SE8900827D0 (en) 1989-03-09
HUT49564A (en) 1989-10-30
AT398073B (en) 1994-09-26
NL8900573A (en) 1989-10-02
GB2216522B (en) 1992-01-02
FR2628419A1 (en) 1989-09-15
ATA50989A (en) 1994-01-15
JPH024746A (en) 1990-01-09
KR890014451A (en) 1989-10-23
GB8905384D0 (en) 1989-04-19
GB2216522A (en) 1989-10-11
BE1003663A3 (en) 1992-05-19
FI93949C (en) 1995-06-26
FI891144A0 (en) 1989-03-09
JP2751068B2 (en) 1998-05-18
HU199775B (en) 1990-03-28
CH678851A5 (en) 1991-11-15
SE8900827L (en) 1989-09-10
FR2628419B1 (en) 1992-02-14
DE3907688A1 (en) 1989-09-21
SE508603C2 (en) 1998-10-19
LU87470A1 (en) 1990-10-02
ES2010439A6 (en) 1989-11-01
IT1229563B (en) 1991-09-04
KR0140993B1 (en) 1998-07-01
FI93949B (en) 1995-03-15

Similar Documents

Publication Publication Date Title
CA1334576C (en) Antiviral pharmaceutical compositions and process for preparing same
CA1339147C (en) Salt-type compounds, pharmaceutical compositions containing them and process for preparing same
GB1591229A (en) Substituted acyl derivatives of amino acids
JPS5912650B2 (en) Process for producing salts of phenyl aliphatic saturated carboxylic acids
GB2048268A (en) Amides of acyl-carnitines process for preparing same and pharmaceutical compositions containing such amides
US4426391A (en) [(Alkoxycarbonyl)oxy]alkyl esters of methyldopa
EP0498268A2 (en) 5-Oxo-L-proline derivatives and pharmaceutical use thereof
GB2116542A (en) Tetrahydroisoquinoline-3- carboxylic acids
PT82134A (en) Process for the preparation of amino acid derivatives comprising the culturing of fr-900490 producing strain belonging to the genus discosia
RU95100773A (en) New esters of sulfuric acid and sugar alcohols
JPH0559105B2 (en)
US4806568A (en) Gossypol derivatives
JPS641474B2 (en)
Anderson et al. [49] Preparation and use of glutathione monoesters
FR2680508A1 (en) NOVEL AMIDE COMPOUNDS OF 1- (ALCOXYBENZYL) PIPERAZINES, PROCESSES FOR PREPARING THEM AND PHARMACEUTICAL COMPOSITIONS CONTAINING SAME
FR2553663A1 (en) 1,2-DITHIOLAN DERIVATIVES, THEIR PREPARATION AND THEIR USE AS MEDICAMENTS
US4407825A (en) Novel bis- and poly-disulfides having immunostimulant activity
FR2622195A1 (en) NOVEL ISOPOLYPEPTIDES, PROCESS FOR THEIR PREPARATION AND MEDICAMENTS CONTAINING SAME
JPH07507786A (en) Use of fatty acids such as Z-hydroxymystic acid as antiviral agents
US3897558A (en) Hypolipemiant and vasodilatatory methods of use
EP0198508B1 (en) L-phosphoserine salts, a process for the preparation thereof and pharmaceutical compositions containing them
EP0032708B1 (en) Tryptophan derivative, process for preparing same and a pharmaceutical composition containing such derivative
CA1138776A (en) Process for producing d-penicillamine and preparation containing same
GB2052985A (en) Tryptophan for use in treating atherosclerosis
JPH0543524A (en) Choline ester of highly unsaturated fatty acid

Legal Events

Date Code Title Description
MKLA Lapsed