JPH02138212A - Treating agent for human immunodeficiency viral disease - Google Patents

Abstract

PURPOSE:To obtain a treating agent for human immunodeficiency viral diseases, containing partially novel specific porphyrins as an active ingredient and having powerful antihuman immunodeficiency viral action with high safety. CONSTITUTION:A treating agent, containing partially novel porphyrins expressed by the formula (R1 to R8 are H, lower alkyl, lower alkenyl, hydroxy lower alkyl, carboxy or carboxy lower alkyl; R9 is H or carboxy lower alkyl, provided that at least one of R1 to R9 is hydroxy lower alkyl, carboxy or carboxy lower alkyl; R10 is H or lower alkyl or R6 and R9 together represent COOCH2; symbols consisting of the solid line and dotted line are single or double bond), N-alkyl substitution product, metal coordination substance, a physiologically acceptable hydrolyzable ester or physiologically acceptable hydrolyzable salt thereof as an active ingredient, having the afore-mentioned effects and effective against the above-mentioned diseases including AIDS, AIDS-related syndromes, persistent general lymphadenitis and lymphoadenopathy.

Description

[Detailed Description of the Invention] [Industrial Application Field] This invention relates to human immunodeficiency virus (Human[n+
The present invention relates to agents for treating various diseases caused by n+undeficiency virus (hereinafter abbreviated as HIV).

[Prior art] HIV is AIDS (acquired immunodeficiency syndrome, AID).
S, Acquired I +u+unodefi
It is known as the virus that causes the virus. AIDS is a viral disease in which T4 lymphocytes are attacked and the immune system is directly destroyed. As a result of the destruction of the immune system, opportunistic infections such as Pneumocystis carinii pneumonia and candidiasis, malignant tumors such as Kaboji's sarcoma, and mental disorders appear, and once AIDS develops, it can last for six months or more unless effective treatment is taken. It is feared as a disease with a poor prognosis, as it can lead to death within a short period of time, such as one year.

If an AIDS epidemic were to occur, it would become a major public health problem because it would cause social unrest and inconvenient situations such as discrimination against patients. Therefore, the development of therapeutic drugs for AIDS is currently an extremely important and urgent issue.

Currently, drugs known to have antiviral effects against HIV include nucleic acid antiviral agents such as azidothymidine (AZT) and special plant ingredients such as glycyrrhizin (licorice component).

[Problems to be Solved by the Invention] However, nucleic acid antiviral agents such as AZT are highly toxic, and plant components such as glycyrrhizin are not sufficiently effective. Therefore, it has been desired to develop an anti-HIV agent that is sufficiently effective and has few side effects.

[Means for Solving the Problem] As a result of searching for substances that have anti-HIV effects among highly safe substances, the inventor found that they were already commercially available as drugs for treating liver diseases, and therefore their safety was confirmed. ing,
It was discovered that protoporphyrin nonodium has a strong anti-HIV effect, and that its analogs also have a similar effect, and based on these findings, the present invention was completed.

That is, this invention relates to the general formula [where R4-R8 is hydrogen, lower alkyl, lower alkenyl, hydroxy lower alkyl, carboxy or carboxyl lower alkyl, and R is hydrogen or carboxy lower alkyl (provided that R, -R, at least one of hydroxy-lower alkyl, carboxy or carboxy-lower alkyl) and Rlo is hydrogen or lower alkyl,
or R8 and R1° together represent -CO-CH,- and R1-R5, R7, R8 and R9 have the above meanings,
A symbol consisting of a solid line and a dotted line means a single bond or a double bond] porphyrins, their N-lower alkyl substituted products, metal ligands, or physiologically acceptable and hydrolyzable esters thereof; The present invention provides an agent for treating HIV disease that contains a physiologically acceptable salt as an active ingredient.

[Embodiments] It is well known that porphyrins exist in tautomeric forms. The above structural formula is shown as a representative of tautomers for convenience, and the present invention includes all tautomers.

The term lower is used to denote groups having 6 or fewer carbon atoms, unless otherwise specified.

In the above formula, lower alkyl includes alkyl groups having 1 to 6 carbon atoms such as methyl, ethyl, propyl, isopropyl, butyl, pentyl, and hexyl.

Lower alkenyl includes groups having 2 to 6 carbon atoms such as vinyl, allyl, l-propenyl, butenyl, pentenyl, hexenyl and the like.

The hydroxy lower alkyl group includes a group having one or more, preferably three or less, hydroxy groups at any position of the above-mentioned lower alkyl group.

Carboxy lower alkyl includes a group having one or more, but preferably less than three, carboxy groups at any position of the above-mentioned lower alkyl group.

Preferred compounds include formula (1) in which R+ Re is the same or different and each represents hydrogen, methyl, ethyl,
vinyl, 1-hydroxyethyl, 3-hydroxypropyl, carboxy, carboxymethyl or 2-carboxyethyl, R8 is hydrogen or carboxymethyl (however,
At least one of these is 1-hydroxyethyl, 3-
hydroxypropyl, carboxy, carboxymethyl or dicarboxyethyl) and RIG is hydrogen or methyl, or R8 and RIQ together -
C(ICH, -1R+ Rs, R7, R8 and R8
is a compound having the above meaning and the symbol consisting of a solid line and a dotted line representing a single bond or a double bond, an N-methyl derivative thereof, or a physiologically acceptable salt thereof.

More preferred compounds are those of formula (I) in which R is methyl, carboxy or carboxymethyl, R3 is hydrogen, methyl, ethyl, vinyl, 1-hydroxyethyl or 2-carboxyethyl, and R3 is methyl, ethyl or carboxymethyl. or 2-carboxyethyl, R4 is hydrogen, methyl, ethyl, vinyl, 1-hydroxyethyl, carboxymethyl or 2-carboxyethyl, R5 is methyl, vinyl, carboxymethyl or 2-carboxyethyl, R8 is hydrogen, methyl, 3 -hydroxypropyl, carboxy, carboxymethyl or 2-carboxyethyl, R7 or methyl, 3-hydroquinepropyl, carboxymethyl or 2-carboxyethyl, Rs is methyl, carboxymethyl or 2-carboxyethyl, RIl is hydrogen or carboxymethyl , R8゜ is hydrogen or methyl, or R11 and R1゜ taken together are 100 GHz-1 RR5, R7, R8 and R
8 has the above meaning.

Examples of typical compounds included in the above general formula are as follows. (The symbol consisting of a solid line and a dotted line indicates a single bond in the case of a chlorophyllin ligand, and a double bond in other cases.) Table 1 Luphyrin 1lle HMe HMe Ce H Ce Me ! 1 Hematoporphyrin Me He Me He Me Ce HCe Me
H Mesoporphyrin Me Et Me Et Me Ce HCe M
e H Philoporphyrin Me Et Me Et Me HMe Ce Me
H protoporphyrin Me Vn Me Vn Me Ce HCe Me
H Fillin Me Et Me Et Me HHCe Me
H Rhodovorufiline Me Et Me Et Me Ca HCe
Me H Uropol” rV Ci+ Ce C+* Ce C
e Ca+ HCe Ca+ H phytoporphyrin Me Et Me Et Me-Co-
Ct(*-Ce Me H chlorophyllin ligand Ca Me El Me Vn Me
It Me Ce Cm Note) M e-methyl, Et
-ethyl, 1-(e-1-hydroxyethyl, Hp=3
-Hydroxypropyl, Vn-vinyl, Ca-carboxy, Cm-carboxymethyl, Ce2-carboxyethyl.

The porphyrins used in this invention are preferably naturally occurring isomers, but mixtures containing these isomers can also be used.

As the metal coordinate of the above compound, Mg is used as the central atom.
1Fe (IIX e.g. heme), Fe(III), FeC
12 (for example, hemin), Co5Cu, and the like are included.

The compounds of the above general formula and their coordinates include known compounds and novel compounds. New compounds can be synthesized in the same manner as known compounds.

Since the above-mentioned porphyrins are amphoteric substances, their physiologically acceptable salts include salts with acids and salts with bases. Physiologically acceptable salts mean salts that do not exhibit significant toxicity at the dosage level and do not have physical or chemical properties that make administration difficult. Among these salts, salts with acids include salts with inorganic acids (mineral acids) such as hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, tartaric acid, citric acid, succinic acid,
Included are salts with organic acids such as carboxylic acids such as fumaric acid and sulfonic acids such as methanesulfonic acid, ethanesulfonic acid, and toluenesulfonic acid. As a salt with a base,
Alkali metal salts such as sodium salts and potassium salts, alkaline earth metal salts such as calcium salts and magnesium salts, polyvalent metal salts such as aluminum salts and manganese salts, trimethylamine salts, triethylamine salts, cyclohexylamine salts, ethanolamine salts, Organic amine salts such as jetanolamine salt, tromethamine salt, morpholine salt, piperidine salt, procaine salt, and caffeine salt are included. When a large number of carboxy groups are present, any number of them can be a salt.

Physiologically acceptable and hydrolyzable esters are esters that do not exhibit significant toxicity at the dosage level and do not have physical or chemical properties that make administration difficult, and that do not occur under physiological conditions, e.g. in the animal body. refers to an ester that can be hydrolyzed under conditions of to give the corresponding acid and alcohol that does not exhibit significant toxicity in the amounts produced. Such esters include lower alkyl esters (e.g. methyl ester, ethyl ester, propyl ester, isopropyl ester, L-butyl ester), lower alkanoyloxy lower alkyl esters (e.g. acetoxymethyl ester, pivaloyloxymethyl ester), l-(lower alkoxycarponyloquine) lower alkyl ester (e.g. l-(ethoxycarbonyloxy)ethyl ester), phthalidyl ester, 5-indanyl ester, 2-(3phthalidylidene)ethyl ester, (5-methyl 2- Oxo-1,3-dioxol-4-yl) methyl ester. If a large number of carboxy groups are present, any number of them can be esters.

HIV is a lymphadenopathy-associated virus (Ly+*pha
Denopathy As5ociated Virus
s+ LAV), human T lymphocyte-tropic virus [1(
Huw+an T-L ymphotropic V
irus Type m: HTLV-I[1], AIDS-associated retrovirus (AID S-As5oc)
iated Retrovirus: ARV)
Although there are mutant strains with different properties such as infectivity to cultured cells, all of these are included.

HIV virus diseases include AIDS (AIDS),
AIDS-related syndrome (ARC), persistent generalized lymphadenopathy (
PGL), and Lymph Juazotubasy syndrome (
According to another classification, acute symptoms due to IV infection, systemic lymph gland swelling, persistent fever,
These include diarrhea, weight loss, dementia, medullary dysfunction, peripheral neuropathy, carinii pneumonia, toxoplasmosis, candidiasis, herpes virus infection, Kabono sarcoma, non-Hodgkin's lymphoma, and cancer.

Treatment includes prevention of infection, prevention of disease onset if infected but not sick, prevention of worsening of disease (maintenance), alleviation of symptoms,
This includes all prevention and treatment, including asymptomatic elimination and complete recovery.

The dosage of porphyrin or its salt for performing the above treatment varies depending on the age, condition, and target treatment of the patient, but is usually 10 to 1000 uy, preferably 5 to 500 uy.
0 to 500 sites, and it is suitable to administer this in divided doses 2 to 4 times a day or as a sustained-release preparation.

For administration, drugs can be administered orally, rectally, externally,
They can be administered in the form of conventional pharmaceutical formulations, mixed with pharmaceutical carriers such as organic or inorganic solid or liquid excipients suitable for administration such as injection.

Such formulations include solids such as tablets, granules, powders, capsules, and liquids such as solutions, suspensions, emulsions, skewers and ointments.

As the carrier, starch, lactose, glucose, sucrose, dextrin, cellulose and its derivatives, paraffin, fatty acid glyceride, water, alcohol, etc. are used.

Also, if necessary, auxiliaries, stabilizers, wetting agents, emulsifiers, lubricants, binders and other conventional additives can be added.

Many of the porphyrins, metal ligands thereof, or salts thereof used in this invention are known to have toxicity, but the toxicity is extremely low. - As an example, the values of protoporphyrin disodium salt are as follows.

Table 2 [Examples] Hereinafter, the present invention will be explained by examples, and the effects of the invention will be clarified by test examples.

In each example, the term "active ingredient" refers to any one of porphyrins, metal ligands thereof, or salts thereof.

Rat 240.3m9/kg Rabbit 220. Oyg/kg Oral administration Mouse >25,000.0m9/kg
Example I Active ingredients 20 mg microscopic cellulose 2519 hydroquinepropyl cellulose 1 m9 carboxymethyl cellulose 5 mg calcium magnesium stearate 2 places lactose
In addition to the appropriate amount, make it a total of 1503F9.

The above ingredients are mixed, granulated by a wet method, compressed into tablets, and coated with sugar by a conventional method.

Example 2 Active ingredient 50R9 physiological saline Add appropriate amount to make a total of 10 mQ.

Test Example 1 (Preparation of sample) The sample was RPMI-1 supplemented with 10% fetal calf serum (FCS).
It was diluted to a predetermined degree at 1640 ml.

(Assay of anti-1-11 V activity) MT-4 cells (MT-4) were infected with the LAV strain of HIV (lO!
・7TCID5o/l11 a+oi O,001T
C.I.D. /cell (or moi O,01TCIDs
o/cell, or moi O,I TCID5o/cell), in the presence of the drug after infection with an adsorption time of 1 h.
After culturing until cytopathic effect (CPE) appeared in the control (no drug added), the amount of HIV produced was measured. The measurements were carried out as follows. M in microplate (96 wells)
T-4 was inoculated at 2x IQ'10.1 mQ/well, and then the virus solution obtained above was inoculated at 0,1 m (1
/ well, observe the appearance of CPE for 7 days, and HI
V production was adjusted to 50% tissue culture infectious concentration (50% Li5s
ue culture 1nfectius dose
: T CI Dso/mc).

Anti-HIV activity was determined when the amount of HIV produced was suppressed compared to the control.

As a drug, protoporphyrin disodium (PP
Dextran sulfate sodium (DSNa) and glycyrrhizin (GR) were used as reference drugs.
and AZT were used.

(Results) The results are shown in the table below, and the numbers in the table are TCID5
o/m (! indicates.

Also, TOX is cytotoxic vinegar.

moi O,I PPMe SNa R ZT moi 0.01 PPMe SNa R ZT Control Table 3 Drug concentration (μg/mc) +03.5 103.0 106.0 Tox + 03.3 102.7 106.0 TOX 108. 0 108・0 106・0 10'・0 <l 01・5 1 O8・0 10'・0 106・0 <101・5 10@・0 moi O,001 PPMe SNa R ZT 102・5 10°・0 10'.0 l 08.0 10".3 l 05.7 Tox <10'.5 Test Example 2 Hemin (HM), hematoporphyrin monosodium (HPNa), protoporphyrin dimethyl ester (PPMe), protoporphyrin as drugs Reductant (first
Table, PP0H), cobroporphyrin III tetramethyl ester (CP lllMe), cobroporphyrin III tetrasodium (CP lllNa), uroporphyrin l dihydrochloride (UPIHCI), deuteroborphyrin dihydrochloride (DP HCI) ), mesoporphyrin dihydrochloride (MPHCI), filoporphyrin (P
hi), pyroporphyrin (P yP ), phytoporphyrin (Pht), N-methylmesoporphyrin” (
MeMP), N-methylprotoporphyrin' (MeMP), N-methylprotoporphyrin' (MeMP),
P P ), chlorophyllin ligand (Chl), and chlorophyll (Ch) were used to test the antiviral activity in the same manner as in Test Example 1 (* indicates an isomer mixture).

The results are shown in the table below.

Table 4 Drug concentration (μg/lll12) +0 1 moio, I HM I HPNa I PPMe l PPOHI CPIIIMe I CP l1lNa IUP I H
CI ID P HCI
IMPHCI Ph1 IPyP
I Pht 104・8 1
0@04・0 l 08 04・t 108 04・2 l08 04・3 108 0′・3 l 08 04・″ 108 04・'tof'04・' +08 04・2 106 04・" 106 04・' 108 MeMP ePP Ch1 moio, 01)-I M P PPMe POH CPIIIMe CPIllNa PHCI PP Chl yP ht Chl h 10'・2 104・! 10'・2 10" ■ 04.0 103.8 104.0 104.0 10'・Violet 10'・1 104.0 10'・0 10'・0 10'・0 10'・0 t 04.0 10'・0 105・0 10"10' O6 M HP PMe POH CpH1Me CP 1llNa PHCI P P hl yP ht h1 103・2 103・0 103・8 103・3 103・3 10'・2 103・2 103・2 103.2 103.2 103.2! 08 10"10" 10@1O1+ OB 10' Describes the inactivation of viruses consisting of irradiating a protein composition with light of separated wavelengths in the presence of a photosensitizer (including, by way of example, porphyrins). Therefore, (1) whether photosensitizers other than porphyrins have anti-HIV activity or not, and (2) whether the porphyrins of this invention are effective against viruses other than HIV, such as herpes simplex virus (H
erpes Symplex V 1rus,
The following experiment was conducted to determine whether the virus has antiviral activity against HSV (abbreviated as HSV).

(1) Anti-HIV activity test of photosensitizers other than porphyrins Antiviral activity was investigated in the same manner as in Test Example 1 using methylene blue (MB), quinoline yellow (QY) and toluidine blue (TB) as test compounds. Ta. The results are shown in the table below.

Test Example 3 Publication No. 275228 describes Table 5 for treatment drug concentration (10μg/mc) moiO,ol MB 10@QY
I O'TB
I Q' From the above results, these are anti-HI
It was found that it had no V activity.

(2) Anti-HIV activity test of porphyrins (cells and viruses) HSVI type WT-51 strain and African green kidney cell line B5-C-40 were used.

H8V stocks were made using B5-C-40, and viral infectivity was measured on B5-C40 monolayers by plaque assay. Eagle's minimal essential medium (MEM) supplemented with 8% neonatal bovine serum was used as the medium, and the cells were incubated at warm C.
O, 3 in incubator (5% COt, 95% air)
Cultured at 7°C.

(Antiviral activity of protoporphyrin) BS-C-4
0 monolayer, I-I S■ with moio, 01 at 37°C.
After infection for 1 hour and washing with MEM, fresh medium was added. After 18 hours of incubation in the presence or absence of protoporphyrin sodium (PPNa), viral infection of the cultures was determined by plaque assay.

(Plaque assay) 0.2 mQ each of serially diluted 10-fold virus solution was
511j! A full monolayer of cells was plated in a plastic Petri dish. After virus adsorption for 1 hour at 37°C, 1%
MEM containing methylcellulose and 2% neonatal bovine serum was overlaid. After 3 days, monolayers were stained with neutral red and plaques were counted.

The results showed that PPNa has no anti-HSV activity.

From the above experimental results, it was found that the therapeutic agent of the present invention has antiviral activity against I(IV).

Claims (2)

[Claims] (1) General formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ [In the formula, R_1 to R_8 are hydrogen, lower alkyl, lower alkenyl, hydroxy lower alkyl, carboxy or carboxy lower alkyl, R_9 is hydrogen or carboxy lower alkyl (However, , at least one of R_1 to R_8
R_1_0 is hydrogen or lower alkyl, or R_6 and R_1_0 together represent -CO-CH_2- R_1 to R_5, R_7,
R_8 and R_9 have the above-mentioned meanings, and the symbol consisting of a solid line and a dotted line means a single bond or a double bond], their N-lower alkyl substituted products, metal ligands, or their physiologically A treatment for human immunodeficiency virus disease, which contains an acceptable hydrolyzable ester or physiologically acceptable salt as an active ingredient. (2) The disease is AIDS (AIDS), AIDS-related syndrome (
2. The therapeutic agent according to claim 1, which is ARC), persistent generalized lymphadenopathy (PGL), or lymphadenopathy syndrome (LAS).