WO2021256547A1

WO2021256547A1 - Composition for treating, preventing, or alleviating sars-cov-2 infectious disease

Info

Publication number: WO2021256547A1
Application number: PCT/JP2021/023116
Authority: WO
Inventors: 厚今泉; 正橋本; 秀昭掛谷; 雅史金井; 幸一渡士
Original assignee: 株式会社セラバイオファーマ; 国立大学法人京都大学; 国立感染症研究所長が代表する日本国
Priority date: 2020-06-19
Filing date: 2021-06-17
Publication date: 2021-12-23
Also published as: JP2023110110A

Abstract

The present invention provides a composition for treating, preventing, or alleviating SARS-CoV-2 infectious disease.　The present invention is: a composition for treating, preventing, or alleviating SARS-CoV-2 infectious disease, the composition containing a curcumin as an active ingredient; and a therapeutic agent.

Description

Composition for the treatment, prevention or alleviation of SARS-CoV-2 infection

The present invention relates to a composition for treating, preventing or alleviating a SARS-CoV-2 infection.

The new coronavirus infection (COVID-19) is an infectious disease caused by the invasion of the new coronavirus SARS-CoV-2 into human cells, which was confirmed in Wuhan City, Hubei Province, People's Republic of China in December 2019. be. As of April 2020, the number of infected people (deaths) has exceeded about 2.5 million cases (about 170,000 cases) worldwide, and has spread to more than 200 countries. The main symptoms are fever, cough, acute respiratory symptoms other than cough, pneumonia, and when it becomes severe, serious organ damage "cytokine storm syndrome" based on excessive immune reaction and severe respiratory failure "acute respiratory distress syndrome" (ARDS) ”can be triggered and fatal.
Since SARS-CoV-2 is an emerging virus, there are no approved drugs at home or abroad. For the treatment of SARS-CoV-2 infection, therapeutic candidate drugs that are diverted from approved antiviral drugs such as remdesivir, nelfinavir, and favipiravir have been developed.
Remdesivir is an antiviral drug developed as a treatment for Ebola hemorrhagic fever. For this remdesivir, the National Institute of Allergy and Infectious Diseases (NIAID) announced on April 29, 2020, as a result of preliminary analysis of clinical trials, that the time to recovery from COVID-19 was significantly longer than that of placebo. It has been announced that it has been shortened.
Nelfinavir is an antiviral drug developed as a treatment for HIV. As a result of non-clinical studies, it has been reported that this nelfinavir exhibits anti-SARS-CoV-2 activity and that the combined effect of nelfinavir and the leukopenia therapeutic agent cepharanthin is observed.
Favipiravir is an antiviral drug that inhibits RNA polymerase, a gene-replicating enzyme of influenza virus. It is expected that it may also be effective against the RNA virus SARS-CoV-2.
In addition, anti-IL antibody drugs having a cytokine interleukin-6 (IL-6) inhibitory action and cytokines are candidates for therapeutic agents for cytokine storm syndrome and acute respiratory distress syndrome in which SARS-CoV-2 infection has become severe. There are Janus kinase (JAK) inhibitors that bind to the receptor.

In recent years, curcumin has pharmacological actions such as tumor formation inhibitory action, antioxidant action, anti-inflammatory action, cholesterol lowering action, antiallergic action (anti-IL-6 action, etc.), brain disease preventive action, and heart disease preventive and therapeutic action. (Patent Document 1).
Also, Mazumar A. Et al. Report on the HIV inhibitory effect of curcumin (Non-Patent Document 1).
However, the effect of curcumin on the new coronavirus infection (COVID-19), the effect of inhibiting SARS-CoV-2 infection, let alone the treatment, prevention or alleviation of SARS-CoV-2 infection by the composition containing curcumin as an active ingredient. There is no knowledge about the effect.
On the other hand, a pharmaceutical composition for parenteral administration, which comprises curcumin as a conjugate with a water-soluble substance (for example, glucuronic acid, sulfuric acid, glutathione, amino acid, etc.) and containing this conjugate as an active ingredient, provides absorption of sparingly water-soluble curcumin. As a result of the improvement, it has been reported that the pharmacological action such as the antitumor action of curcumin can be sufficiently obtained (Patent Document 2).

Japanese Unexamined Patent Publication No. 2013-95723 WO2018 / 003857 Gazette

As described above, antiviral drugs such as remdesivir, nerfinavir, and favipiravir have been developed as therapeutic drug candidates for SARS-CoV-2 infection, but the effect as a therapeutic drug is improved and the degree of freedom in selecting the therapeutic drug is ensured. In addition to these, new therapeutic agents for SARS-CoV-2 infection are required from the viewpoint of reducing side effects.
That is, it is an object of the present invention to provide a novel therapeutic composition for SARS-CoV-2 infection.

Therefore, the present inventors investigated the SARS-CoV-2 infection inhibitory action of various components, and found that curcumins exhibited an excellent SARS-CoV-2 infection inhibitory effect, and completed the present invention.

That is, the present invention includes the following.
[1] A composition for treating, preventing or alleviating SARS-CoV-2 infection, which contains curcumin as an active ingredient.
[2] The composition according to the above [1], wherein the curcumins are one or more selected from curcumin, curcumin derivatives, and conjugates thereof with a water-soluble substance.
[3] The composition according to the above [2], wherein the curcumin derivative is at least one selected from bisdemethoxycurcumin, demethoxycurcumin and tetrahydrocurcumin.
[4] The composition according to the above [2], wherein the water-soluble substance is at least one selected from glucuronic acid, sulfuric acid, glutathione and amino acids.
[5] The composition according to the above [2], wherein the water-soluble substance is at least one selected from glucuronic acid and sulfuric acid.
[6] The composition according to the above [1], wherein the curcumins are curcumin monoglucuronide.
[7] The composition according to any one of the above [1] to [6], wherein the SARS-CoV-2 infection is a cytokine storm syndrome or an acute respiratory distress syndrome.
[8] The composition according to any one of the above [1] to [7], which is for parenteral administration.
[9] Curcumins for use in the treatment, prevention or alleviation of SARS-CoV-2 infections.
[10] The curcumins according to the above [9], wherein the curcumins are curcumin monoglucuronides.
[11] Use of curcumins for the production of compositions for the treatment, prevention or alleviation of SARS-CoV-2 infections.
[12] The use according to the above [11], wherein the curcumins are curcumin monoglucuronides.
[13] A method for treating, preventing or alleviating a SARS-CoV-2 infection, which comprises administering an effective amount of curcumin.
[14] The method according to the above [13], wherein the curcumins are curcumin monoglucuronide.

The composition for treating, preventing or alleviating SARS-CoV-2 infection containing the curcumins of the present invention as an active ingredient has side effects due to its excellent anti-SARS-CoV-2 activity, as shown in Examples below. A reduced therapeutic, preventive or palliative effect on SARS-CoV-2 infection can be obtained.
As a result of the expression of the antiviral effect of curcumins, and due to the antiallergic effect of curcumins, it can be expected to have an inhibitory effect on cytokine storm syndrome or acute respiratory distress syndrome due to the aggravation of SARS-CoV-2 infection. (Non-Patent Document 2).
The composition of the present invention can be used, for example, in pharmaceutical compositions, supplements, functional foods, etc., without limitation in its use.

It is a figure which shows the result of the SARS-CoV-2 infection inhibitory effect of Example 1. FIG.

The active ingredient of the composition for treating, preventing or alleviating the SARS-CoV-2 infection of the present invention is curcumin.
Examples of the curcumins include one or more selected from curcumin, curcumin derivatives, and conjugates thereof with a water-soluble substance.

Curcumin is the main component of curcuminoids contained in turmeric pigments, and is a compound represented by the following structural formula (1).

Derivatives of curcumin include bisdemethoxycurcumin, demethoxycurcumin and tetrahydrocurcumin.
In the present invention, as curcumin, chemically synthesized curcumin may be used, or curcumin distributed as a turmeric pigment may be used. As the turmeric pigment, turmeric powder obtained by powdering dried rhizome of Curcuma longa LINNE, and the turmeric powder using an appropriate solvent (for example, ethanol, fat, propylene glycol, hexane, acetone, etc.) are used. Examples thereof include crude curcumin or oleoresin (turmeric oleoresin) obtained by extraction, and purified curcumin.
In addition, curcumin includes both keto type and enol type which are tautomers.

Examples of the water-soluble substance in the conjugate of the curcumin or curcumin derivative of the present invention and the water-soluble substance include one or more selected from glucuronic acid, sulfuric acid, glutathione and amino acids. Examples of amino acids include amino acids existing in the living body, for example, essential amino acids.
For example, a conjugate of curcumin and a water-soluble substance can maintain a high blood concentration of free curcumin. As a result, the pharmacological action of curcumin can be sufficiently obtained. Further, the conjugate of curcumin and a water-soluble substance is preferable because it is a metabolite of curcumin in vivo and has very high safety.
The binding molar ratio of curcumin to the water-soluble substance is preferably curcumin: water-soluble substance = 1: 1 to 1: 3, more preferably 1: 1 to 1: 2, and even more preferably 1: 1.

The conjugation form (binding form) of curcumin and the water-soluble substance is, for example, the form of the formula (2).

(In the formula, ^{at least one of R 1} and R ² is a residue of the water-soluble substance, and the remainder is a hydrogen atom.)

In the formula (2), ^{one or both of R 1} and R ² are preferably glucuronic acid residues or sulfuric acid residues, and the remainder is preferably a hydrogen atom. Of these, curcumin monoglucuronide, in which R ¹ is a glucuronic acid residue and R ² is a hydrogen atom, is more preferable.
The conjugate of curcumin and a water-soluble substance can be produced by the method described in Patent Document 2.

Examples of the SARS-CoV-2 infectious disease targeted by the composition of the present invention include cytokine storm syndrome and acute respiratory distress syndrome.

The content of the water-soluble substance conjugate of curcumins in the composition of the present invention is the content of the SARS-CoV-2 infection, including the case where it is used as a therapeutic agent for the treatment, prevention or alleviation of the SARS-CoV-2 infection. Since it is appropriately determined according to the type, gender, age, symptom, etc. of the patient, it cannot be unconditionally determined, but for example, 1 to 100% by mass is preferable, 5 to 100% by mass is more preferable, and 10 to 100% is more preferable. %% by mass is more preferred.

The composition of the present invention is not particularly limited as a dosage form when used in the form of a therapeutic agent. For example, it can be used as a liquid preparation or a lyophilized preparation prepared at the time of use. It may also be a sustained release dosage form or a sustained release dosage form. It can also be prepared by using additives such as carriers and excipients usually used for the preparation.
The administration route of the therapeutic agent of the present invention can be selected from systemic administration and local administration, oral route and parenteral route according to the disease, symptom, etc., and is a suitable agent depending on the administration method and route. Oral administration in the form of tablets, rounds, capsules, granules, powders, solutions, etc., or injections (eg, intravenous, intramuscular, etc.), suppositories, transdermal agents, nasal agents, etc. Parenteral administration in the form of an inhalant or the like can be selected.

The therapeutic agent for oral administration according to the present invention may be a solid preparation such as tablets, capsules, powders and granules. Such a preparation is produced according to a conventional method by mixing one or more active substances with an inert excipient, lubricant, disintegrant, solubilizing agent and the like. Excipients can be, for example, lactose, cellulose, mannitol, glucose. The lubricant can be, for example, magnesium stearate. The disintegrant can be, for example, sodium carboxymethyl starch. Tablets or pills may be coated with a sugar coating or a gastric or enteric coating, if desired.

The therapeutic agent for oral administration may be a pharmaceutically acceptable liquid preparation such as an extract, an emulsion, a liquid, a suspension, a syrup, a liquor, or an elixir. Such formulations contain commonly used inert solvents (eg, purified water, ethanol, etc.) and further include solubilizers, wetting agents, suspending agents, sweeteners, citrates, fragrances, etc. It may contain a buffer (eg, sodium citrate, etc.), a stabilizer or a preservative.

Therapeutic agents for parenteral administration include sterile aqueous or non-aqueous liquids, suspensions, or injections such as emulsions, ointments and lotions, sublingual agents for oral administration, oral patches, and nasal administration. Can be an aerosol or suppository for.
In the case of an injection, it can be administered by injection into the joint, subcutaneous, intradermal, intramuscular, etc., in addition to the usual intravenous administration and intraarterial administration. The aqueous solvent for the injectable solution can be, for example, distilled water or saline. The non-aqueous solvent for the injectable solution can be, for example, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, alcohols such as ethanol, or polysorbate 80 (Public name). Such formulations also include tonicity agents (eg, sodium chloride, glucose, etc.), preservatives, wetting agents, emulsifiers, dispersants, stabilizers, pH adjusters (eg, sodium citrate, sodium acetate, phosphorus). It may contain a buffer (eg, sodium acetate, etc.), a local anesthetic (eg, prokine hydrochloride, lidocaine hydrochloride, etc.) or a solubilizing agent.
These formulations can be sterilized, for example, by filtration through a bacterial retention filter, formulation of a fungicide, or irradiation. Moreover, the composition obtained by dissolving or suspending a sterile solid composition in sterile water or an injection solvent before use can also be used as these preparations. These preparations can be produced by a known method usually used in the preparation process.

When the composition of the present invention is a therapeutic agent, a prophylactic or palliative therapeutic agent, it is preferably administered as an injection. In this case, the form of the composition of the present invention may be a powder filler in which it is dissolved at the time of use, or it may be an aqueous solution. Compositions for parenteral administration include water-soluble substance conjugates of curcumins, water, physiological saline, pH regulators, sugars, acids, alkalis, buffers, isotonic agents, stabilizers, and soothing agents. , Preservatives and the like can be blended.
Examples of saccharides include monosaccharides, disaccharides, trisaccharides, polysaccharides, sugar alcohols and the like. Examples of the acid and alkali include water-soluble inorganic acids, water-soluble inorganic acid salts, water-soluble organic acids, water-soluble organic acid salts, amino acids, amino acid salts and the like.

The composition of the present invention may further contain other pharmaceuticals in combination as part of the active ingredient. Such drugs include drugs for the treatment of SARS-CoV-2 infections; antiviral drugs such as remdecibir, nerfinavir, favivillavir, ropinavir, ritonavir, steroid drugs such as cyclesonide, and antiinflammatory drugs such as chlorokin and hydroxychlorokin. Drugs with antiviral activity of proteolytic enzyme inhibitors such as nafamostat and camostat; for example, anti-IL-6R antibodies such as tocilizumab and salilumab, JAK inhibitors such as baricitinib and luxolitinib, BTK inhibitors such as acarabrutinib, labrizumab and the like. Examples thereof include drugs for the treatment of severe cytokine storm syndrome and acute respiratory distress syndrome such as anti-complementary antibody.
The content of these other drugs is appropriately determined according to the type of SARS-CoV-2 infection, the sex, age, symptoms and the like of the patient. The same applies when used as a pharmaceutical composition for the prevention or alleviation of SARS-CoV-2 infection.

Next, the present invention will be described in more detail with reference to examples, but the scope of the present invention is not limited to the specific aspects described below.

Production Example 1
(Preparation method of a conjugate of curcumin and glucuronic acid, which is a water-soluble substance (curcumin monoglucuronide))
Curcumin monoglucuronide was synthesized according to the method described in WO2018 / 03857. That is, compound (1) (β-D-glucopyranoside uronic acid, 4) was reacted with 1.0 g (2.52 mmol) of acetbromo-α-D-glucuronic acid methyl ester and 326.0 mg (2.15 mmol) of vanillin as raw materials. -Formyl-2-methoxyphenyl, methyl ester, triacetate) 491.6 mg (yield 49%) was obtained. On the other hand, compound (2) (5-hydroxy-1- (4-hydroxy-3-methoxyphenyl) -1, 4-Hexadiene-3-on) was obtained. Deacetylation of the obtained compound (3) (curcumin β-D-glucopyranoside uronic acid 2,3,4-tri-0-acetyl, methyl ester) by reacting the compound (1) with the compound (2). Purification gave curcumin monoglucuronide (253.1 mg).

Example 1
(SARS-CoV-2 infection in vitro inhibitory effect of curcumin)
Matsuyama, S.A. et al. According to PNAS, 117, 7001 (2020) "Enchanted isolation of SARS-CoV-2 by TMPRSS2", the inhibitory effect of curcumin on SARS-CoV-2 infection (in vitro) was confirmed using VeroE6 / TMPRSS2 cells. VeroE6 / TMPRSS2 cells are conserved in (JCRB no. JCRB1819).
(1) TMPRSS2 overexpressing Vero6 cell line was monolayer-cultured in each well of a 96-well cell culture plate manufactured by IWAKI. Then, as a test drug,
The concentrations of the curcumins of the present invention (curcumin monoglucuronide obtained in Production Example 1) were 50 μM, 16.7 μM, 5.56 μM, and 1.85 μM, respectively.
-Remdesivir 10 μM (MedChemExpress) and Nelfinavir 8 μM (Japan Tobacco) as a positive drug control group, and dimethyl sulfoxide 0.5% (Sigma) as a virus control group.
Was added, and the cells were cultured at 37 ° C. for 1 hour under _{5% CO 2 conditions.} DMEM medium manufactured by Life Technologies and fetal bovine serum were used for culturing.
(2) SARS-CoV-2 (obtained from National Institute of Infectious Diseases Virus Part 3) was administered to the cells in each well after the culture at an infection multiplicity of infection MOI = 0.01. Then, after washing to remove excess SARS-CoV-2, the cells were cultured at 37 ° C. for 24 hours in the coexistence of the above drugs and the like.
(3) After completion of culturing, RNA of SARS-CoV-2 in the supernatant of the culture broth of each well was quantified using a real-time PCR system (Applied Biosystems) and combined with reactive viral RNA (relative viral RNA blacked out). Cell virus (white cell viability) was determined.
The cells treated in the same manner without the addition of SARS-CoV-2 were used as a normal cell control group, and the same calculation was performed.
(4) The results are shown in FIG. (In FIG. 1, curcumin is labeled as Curcumin, remdesivir is labeled as remdesivir, nelfinavir is labeled as nelfinavir, dimethyl sulfoxide is labeled as DMSO, and the normal cell control group is labeled as virus (-).)
From FIG. 1, curcumin 50 μM showed a reactive viral RNA inhibitory effect superior to that of the positive drug control group while maintaining the same cell viability, and was calculated as _{EC 50 = 25.2 μM for the TMPRSS2 overexpressing VeroE6 cell line of curcumin.} .. This is better than the _{EC 50 => 64μM (in vitro} ) of favipiravir (https://doi.org/10.1101/2020.04.14.039925.). The EC ₅₀ = 25.2 μM concentration of curcumin is a concentration that can be sufficiently achieved intravenously with curcumin monoglucuronide as in the invention described in Patent Document 2. Therefore, the composition of the present invention is sufficiently effective against SARS-CoV-2 infection.

Claims

A composition for treating, preventing or alleviating SARS-CoV-2 infection, which contains curcumin as an active ingredient.
The composition according to claim 1, wherein the curcumins are one or more selected from curcumin, curcumin derivatives, and a conjugate of them and a water-soluble substance.
The composition according to claim 2, wherein the curcumin derivative is at least one selected from bisdemethoxycurcumin, demethoxycurcumin and tetrahydrocurcumin.
The composition according to claim 2, wherein the water-soluble substance is at least one selected from glucuronic acid, sulfuric acid, glutathione and amino acids.
The composition according to claim 2, wherein the water-soluble substance is at least one selected from glucuronic acid and sulfuric acid.
The composition according to claim 1, wherein the curcumins are curcumin monoglucuronide.
The composition according to any one of claims 1 to 6, wherein the SARS-CoV-2 infection is a cytokine storm syndrome or an acute respiratory distress syndrome.
The composition according to any one of claims 1 to 7, which is for parenteral administration.
Curcumins for use in the treatment, prevention or alleviation of SARS-CoV-2 infections.
The curcumin according to claim 9, wherein the curcumin is curcumin monoglucuronide.
Use of curcumins for the production of compositions for the treatment, prevention or alleviation of SARS-CoV-2 infections.
The use according to claim 11, wherein the curcumins are curcumin monoglucuronides.
A method for treating, preventing or alleviating SARS-CoV-2 infection, which comprises administering an effective amount of curcumin.
13. The method of claim 13, wherein the curcumins are curcumin monoglucuronides.