JPS62501357A - Antiviral and antitumor cyclohexadienone compositions - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Antiviral and antitumor cyclohexadienone compositions (Continuation application data) This application is based on U.S. Patent Application No. 682,278, filed December 17, 1984; This is a partial continuation of No. 744620 filed on June 14, 1985.

(Field of invention) The present invention discloses a novel cyclohexadienone with useful antiviral and antitumor activities. Regarding derivatives. More specifically, the present invention relates to marine organisms derived from red algae. The present invention relates to cyclohegisadienone derivatives having antiviral and antitumor activities.

(Background of the invention) Viral diseases cause suffering to humans, plants, insects and animals.

The prevention and control of viral diseases has important health and economic significance.

Viral diseases are associated with human pain, including the common cold, herpes, and cancer, so it is important to control them. Something important is clear. Economic reasons and spreading viral diseases to humans Prevention of viral diseases in animals against the ability of animals to act as sources or vectors of viruses. Exclusion is also important. Viral diseases of plants are common in fruit trees, tobacco and various vegetable crops. It is known to have a devastating effect on culture. Another person infected with insect virus disease There is interest because of the ability of insects to transmit viral diseases.

In this way, the prevention and control of viral diseases is extremely important for humans, and we Research has been devoted to antiviral methods. inhibit, prevent or destroy viruses Although some methods and chemical compositions have been developed that are part of and antiviral chemical compositions.

Calvin and Ellis U.S. Pat. Nos. 4,162,308 and 4,162,509 has shown that a water-soluble extract from marine red algae inhibits the growth of certain herpes viruses. He says it has been found to be effective.

U.S. Patent No. 4,152,508 discloses that Tsurnerella nortensiana selected from the group consisting of Epiphytic Force, Tsurnerella Benni and mixtures thereof S. red algae that have been infected with type 1 and type 2 herpes simplex virus and herpes zoster virus. It inhibits the proliferation of viruses and reduces the pain caused by infection by those viruses. It is said that it is effective in eliminating

U.S. Patent No. 4,162,308 covers type 1 and type 2 herpes simplex virus and and herpes zoster virus, and prevents infection and infection by these viruses. Neodylusia ψ Americana and A water-soluble extract from a selected group of marine red algae consisting of Neodylusia inchigra. It describes examples of uses for artifacts.

The entire disclosures of U.S. Pat. Nos. 4,162,508 and 4,162,309 are hereby incorporated by reference. I will cite ib as a reference.

Cruise was published in 1984 by Bergamon Press Limited, UK. "L Sentence - Long No. 1" A Volume 28, No. 7.

pp. 1449-1451, ['Biologically active monoterpenes from red algae'] describes the biological activities of monoterpenes collected from Cruise et al. A halogenated monote extracted with methylene chloride from the red algae Kas hornemanni. Discloses le pens. Halogenated monoterpenes are used in various applications. It was shown to be bioactive and biotoxic to insects.

The water-soluble red algae extract described in the Calvin and Ellis U.S. Patent Application No. 1 , eat red algae and red algae in addition to the halogenated monoterpenes of Cruz et al. Other compounds have been isolated from the mollusk known as Aplysia. There is. These compounds include the halogenated chamigranes, which were introduced in 1978. Published by Academic Press, 1983, Marine edited by P. J. Nyoyer. eSujural Products Volume 1 (Martin) and m5s (Ericsson) The full disclosure of this document is incorporated herein by reference. Give it as a gift.

The inventors have also found that extracts of red algae and the red algae-eating Aplysia spp. The company has filed patent applications for compounds that exhibit antiviral activity. Certain halogenated chamigranes, i.e., US patent application filed December 18, 1984. National Patent Application No. 682896 and certain species exhibiting antiviral and antitumor activity cyclohexadienone compounds, i.e., December 17, 1984 and and U.S. Patent Application Nos. 682,278 and 7446, filed June 14, 1985. The entire disclosure contents of these three simultaneous applications, including No. 20, are included here as cited documents. give.

Another simultaneous application filed by the inventors at the same time as this application deals with red algae and red algae. Certain halogens prepared from edible Aplysia extracts and show anti-tumor activity The present invention relates to compounds containing chamigranes.

Tumor prevention, growth control and regression in mammals is also important in humans . Kanashi's research has been devoted to oncology and anti-tumor measures. The term tumor , an abnormal cell mass that is a new tissue growth that disrupts the order of the original tissue or the host's entire body. means. Tumors are common in various mammals, and tumor prevention, growth suppression, and degeneration are important. is important to people.

Tumors are a type 4 form of cancer and associated with various physical disorders including cancer cachexia. KIQ4 causes pain to mammals and humans. Cancer cachexia means having a tumor. Included in mammalian suffering. The fear of cancer is such that, for example, cancer is a major cause of death for humans, including the heart and the heart. It is also well known because it is the second most common disease after vascular and vascular diseases.

Kanashi research and resources have been devoted to oncology and anti-tumor measures, including chemotherapy. . Certain methods and chemical compositions may be part of inhibiting, mitigating, and controlling tumor growth. At the same time as chemical compounds have been developed, new methods and antitumor chemical compositions are required. It is.

Certain cyclohexadienone compounds derived from red algae extracts are useful anti-inflammatory agents. It has now been found to have viral and anti-tumor activity.

(Summary of the invention) It is therefore an object of the present invention to create novel compositions and compositions useful as antiviral and antitumor agents. and a process for producing such novel antiviral and antitumor compositions. There is a particular thing.

Another object of the invention is to use novel antiviral and antitumor compositions, respectively. The objective is to provide a method for inhibiting viruses and tumors.

Further objects and utilities of the invention are set forth in part in the following description and in part in , may be obvious from the description, or may be learned by practice of the invention.

The object and utility of the invention lies in the compositions, processes and methods particularly pointed out in the appended claims. and is realized and achieved by means of combination.

To accomplish the objectives in accordance with the objectives of the invention, embodiments are herein shown and fully described. As stated above, the present invention comprises compositions of the general formula.

Both R groups may be the same or different, hydrogen or a lower alkyl group, and X is a lower alkyl group. alkyl, hydroxy, lower R-y syloxy, lower alkoxy, fluoro, R is hydroxy, lower acyloxy group, bromo or iodo group, It is a halogenated lower acyloxy group or a halogen.

In a preferred embodiment of the present invention, R is a methyl group and R2 is an acetyloxy V group. , and X is methyl, chloroIV or bromo group.

In a further preferred embodiment of the present invention, the present invention provides the formula: consisting of a composition of As shown in embodiments and fully described herein, the present invention comprises as active ingredients Formula I or or preferably an antiviral or antitumor effective amount of one or more of the compositions comprising and an antiviral or antiviral drug consisting of a non-toxic bulking agent or diluent approved by the Pharmacy Act. Consisting of an oncogenic composition.

As particularly shown in embodiments and extensively described, the present invention also provides for the production of red algae. The red algae are collected and contacted with a suitable organic solvent to obtain an extract, and the compound of formula A method of preparing a compound of formula I comprising the step of isolating the compound of formula I. Preferably one The method involves collecting red algae, especially Desmia hornemanni, and treating the red algae with a suitable first species. A derivative of the first compound of formula 1, preferably a derivative of formula 1, is obtained by contacting with an organic solvent. The solvent containing the alcohol derivative is removed by conventional methods, preferably chromatography. The derivative of 2 is isolated by the method, and this alcohol derivative is acetylated to produce 2 It consists of each step to obtain a compound.

As particularly illustrated and fully described in embodiments, the present invention further provides The tumor is treated with one or more antiviral or antitumor compositions of Formula It or preferably Formula H. From a method of inhibiting a virus or tumor comprising contacting with an effective amount Become.

In accordance with the objectives of the invention, one embodiment of the invention is herein shown and fully described. further consists of a composition of the general formula (Seal): ■ The NLR is: CONH2 Here, X is oxygen, X is hydrogen, halogen, lower alkyl group or alkyl sulfate. Phenyl group, sulfur or imine group; Y is oxygen or sulfur; Z is oxygen, sulfur or The imine group and R1 group are the same, hydrogen or lower alkyl group, and R2 is H or lower alkyl group. alkyl group, and n is 1 to 5.

X1 and Y are methyl groups and are oxygen.

In a more preferred embodiment of the invention, the invention provides the formula consisting of a composition of

As shown in embodiments and fully described herein, the invention also provides a right-handed component. In this case, an antitumor composition comprising a material rR effect Zg, patvzp formula I or a diluent is used. Ru.

As shown in embodiments herein and fully described, the present invention also provides methods for harvesting red algae [7]. , the red algae is brought into contact with a suitable organic solvent to obtain an extract, and the second compound is produced. It consists of a method for producing a compound consisting of each step of isolation.

Embodiment L2 herein, as fully described, the present invention further provides anti-tumor efficacy. of one or more of formula I or I, or preferably one of formulas ■, ■, ■ or ■ A method for inhibiting a tumor comprising contacting the tumor with a composition. child As illustrated and fully described herein, the present invention also provides treatment for cancer cachexia conditions. Inhibit, reduce and control the growth of tumors or tumor cells that cause symptoms This consists of several methods of treating cancer cachexia.

The detailed descriptions above and below are merely exemplary and explanatory, and do not apply to claims. It should be understood that no limitations on the invention are intended.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS OF THE INVENTION Here, preferred embodiments of the invention Let me mention it in detail. An example of this is explained in the Examples section below. In accordance with the invention, the formula: A composition is provided.

Tasoshi R1 groups are the same or different, hydrogen or lower alkyl yi, X is lower alkyl Kyl, hydroxy, lower acyloxy, low m alkoxy, fluor, chlor, Promata is a halogenated lower acyl group or a halogen.

Preferably, lower alkyl and acyloxy groups have 1 to 5 carbon atoms, More preferably 1 to 3 carbon atoms, most preferably lower alkyl group is methyl, and the lower acyloxy group is acetyloxy.

Preferably, X is methyl, chloro or bromo group, more preferably X is chloro group It is.

More specifically, a preferred embodiment of the invention comprises a composition of structural formula H: : ■ Another preferred embodiment of the invention consists of a composition of the following formula: I Antiviral compositions comprising legally approved non-toxic bulking agents or diluents are provided. Ru. Although the effective amount changes depending on the usage conditions of the antiviral composition, it is generally The minimum dose required to show activity is between 25 and 806 viral plaque-forming units. 50 to 200 micrograms. Non-toxic, Pharmacy Law approved, non-toxic filler or Examples of useful diluents include, but are not limited to: ethanol. dimethyl sulfoxide and glycerin.

According to the invention, the process for preparing compounds according to formula 1 comprises Runemanni (also known as Chondrocottus hornemanni), collect red Contacting the algae with a suitable organic solvent to obtain a solvent extract and isolate the compound of formula 1 It consists of each process.

Detailed description of preferred embodiments of the process of the invention for preparing compounds of formula 1 The explanation is as follows. Red algae.

Desmia hornemanni was collected at Cape Zampa, Okinawa. Next, acetate this red algae. (first solvent) to obtain an acetone extract from red algae. Next, aceto The extract was concentrated by evaporation under heat or reduced pressure, and the acetone residue was dissolved in methylene chloride. to obtain a methylene chloride extract.

Acetone and methylene chloride are currently the preferred choices for the first and second solvents. However, other suitable solvents may be substituted for acetone and/or methylene chloride. stomach. A suitable first solvent is one that is capable of extracting the compound of formula ■ from other components of the red algae. It must be the. A suitable first solvent that can be used in place of acetone is Examples include, but are not limited to, the following organic solvents: methyl ethyl ketone, ethyl acetate. methanol, ethanol and methyl isobutyl ketone.

A suitable second solvent extracts the compound of formula 1 from other components present in the first solvent extract. It must have the ability to extract and separate. Even if the drink is replaced with methylene chloride Suitable second solvents include, but are not limited to, the following organic solvents: :Chloroform, trichlorethylene, hexane, and lower alkanes. skilled in the art As is known to those skilled in the art, various ratios of first and second solvents may be used in the present invention.

The methylene chloride extract is removed and treated with methylene chloride either by gentle heating or under reduced pressure. Evaporate and concentrate the tyrene solvent.

The alcohol derivative of the compound of formula] is converted into methylene chloride by chromatography method. isolated from the extract.

Methylene chloride extracts may be used in any suitable romatographic technique. was first chromatographed on a silica gel column to create a mixture of hexane and acetone. Alcohol derivatives of formula n were eluted with hexane-acetone in a liter ratio of about 7:3. Obtain the main fraction of the body, and then apply the main fraction to thin-layer chromatography-grade silicone. The compound of the formula is separated by flash chromatography method on Kagel column. It was found that a pure alcohol derivative of the product can be obtained. Other solutions known to those skilled in the art A solvent and a column may be used instead.

The alcohol derivative of the compound of formula l was found to decompose within 2 hours at room temperature. The best thing to do is to carry out the next step as quickly as possible.

The alcohol derivative of the compound of formula I is prepared in a milliliter ratio of acetic anhydride to pyridine of about 1: Acetylate with a mixture of hyridine acetate anhydride of 1 for about 10 minutes. Currently, acetic anhydride A preferred choice of acetylating agent is a mixture of 1, anhydrous In place of acetic acid, impropenyl acetate and acetyl chloride, in place of pyridine, N- With other suitable agents such as methylmorpholine and N-methylpyrrolidine Good too. Excess acetic anhydride and pyridine are removed by evaporative dehydration to obtain the pure formula obtain a compound.

be inhibited from proliferating or killed by methods consisting of exposure. Generally the above The minimum effective dose is 25 to 80 virus bed forming units per 50 to 100 viral bed forming units. It is an ikrogram. Compounds of formulas 1 and 1 can be used to treat RNA viruses, varicella oral Flame (rVSVJ, !: call), arenavirus, coronavirus, influenza virus, and the DNA virus, herpes simplex-1 (rH8V-IJ). b) Includes other herpes viruses, adenoviruses and bapopaviruses, but , has activity to inhibit or kill a wide range of viruses, including but not limited to.

The efficacy of the composition of the invention to inhibit viral cells is thereby inhibited and destroyed. Hostosinfections caused by viruses that include, but are not limited to, virus, coronavirus, reovirus, influenza virus, etc. Those caused by RNA viruses, as well as herpes virus, adenomatous caused by NA viruses such as Virus and Babopavirus Contains things. The present invention is also useful in controlling common viral infections in plants. be.

According to the invention, the formula: A composition is provided.

Taguchi, R is Here, X is oxygen, sulfur or imine group; X is hydrogen, halogen, lower alkyl or alkylsulfenyl group iY is oxygen or sulfur; Z is oxygen, sulfur or is an imine group 1 Both R are the same or different hydrogen or a lower alkyl group %R'' is H or a lower alkyl group; and n is 1 to 5.

Preferably the lower alkyl group has 1 to 5 carbon atoms, more preferably 1 to 5 carbon atoms. Having 3 carbon atoms, the most preferably lower alkyl group is methyl.

Preferably X and Y are oxygen.

More specifically, preferred embodiments of the present invention include the following products: ■, ■, ■, or consisting of a composition of

■v According to the present invention, the active ingredient is a non-toxic bulking agent or provides an anti-tumor composition comprising a diluent. Changes in the conditions of use of antitumor compositions Although the effective dose varies depending on the number of tumor cells, in general, the minimum dose required for activity is O pieces, 1 to 100 micrograms. Non-toxic dosage increase approved by Pharmacy Law Useful examples of agents or diluents include, but are not limited to: Mu: ethanol, dimethyl sulfoxide and glycerol.

According to the present invention, the method for producing a stratified compound is a method for producing a stratified compound using red algae, especially desmia hornemanni. (known as Chondrococcus φ hornemanni), and this red algae was Contact with an organic solvent to obtain a solvent extract, and then synthesize and immediately release compound 2. Consists of each process.

Detailed description of preferred embodiments of the process of the invention for producing compounds of Niso The explanation is as below. Red algae, Desmia hornemanni, at Cape Zamba, Okinawa Collect. Next, this red algae is brought into contact with acetone (first solvent) to remove acetate from the red algae. Obtain ton extract. The acetone extract is then evaporated either by heating or under reduced pressure. Concentrate by dehydration method, bring the acetone residue into contact with methylene chloride, and extract with methylene chloride. get something

Currently, acetone and methylene chloride are the preferred choices as second solvents. However, other suitable solvents can also be used in place of acetone and methylene chloride. A suitable first solvent may be used to extract the compound of formula I from other components of the red algae. must have the ability to Suitable first solvent that may be used in place of acetone includes, but is not limited to, the following polar organic solvents: methyl ethyl keto ethyl acetate, methanol, ethanol, and methyl isobutyl ketone. suitable Suitable second solvents include compounds of formula I from other components likely to be present in the first solvent extract. It must have the ability to extract and separate compounds. Can be used instead of methylene chloride Good suitable second solvents include, but are not limited to, the following organic solvents: :Chloroform, trichlorethylene, hexane and lower alkanes. person skilled in the art As is known in the art, various ratios of first and second solvents may be used in the present invention.

Take out the methylene chloride extract and add methylene chloride to it under gentle heat or reduced pressure. The solvent was concentrated by evaporation and dehydration. Alcohol derivatives of compounds of formula ■ ()1) was isolated from the methylene chloride extract by chromatographic methods.

The alcohol derivative has the following formula: Any suitable chromatography technique You can also use squid, 'f, -f methylene chloride extract in a silica gel column. Chromatograph into 5 ml of hexane to acetone at a ratio of approximately 7:3. Elute with gisan-acetone to obtain the main fraction containing compound 2, and then further dilute it. Flash chromatography on layer chromatography-grade silica gel cuffs It was found that pure formula 111 can be obtained by separating the main fraction using the Fee method. To those skilled in the art Other eluents and columns may be used instead, as is known in the art. The compound of formula 11 is It decomposes within 2 hours at room temperature, allowing the next synthesis step to be carried out as quickly as possible. is important.

Preferred embodiments of the present invention according to the formula ■%V%'%1 and For the purpose of preparing those compounds from compounds of formula 11 and ■ in order to Let us take up the synthesis process described below.

Compounds of formulas ■ and V are of the formula: ■ is synthesized from a halogen derivative of a compound of formula (1) having

Shifter X3 is CI or Br.

The compound of formula ■ can be prepared as described above to obtain the alcohol derivative of formula 1 (formula 1N). The volatile oil was removed from the methylene chloride extract subjected to chromatography, and the By converting the volatile oil into a halogen derivative of the compound of formula I having the formula That's what you get.

The halogen derivative (isolation, e.g. by thin layer chromatography method) After separation by ) a large amount of ketal of formula V and a small amount of vinyl ether of formula (2) are obtained.

The compound of formula OH: Obtained by hydrolyzing the ketal of formula V with 5N HC1).

The compound of formula A base such as chlorine (e.g., 1% KOH aqueous solution nidioxane, l=l by volume) It is inhibited by a method in which it is synthesized by treatment with In general, the minimum effective dose as described above is Each number lθ is 1 to 100 micrograms. Compounds of formulas I to ■ are Pulmonary carcinoma A349゜ileocecal adenocarcinoma HCT-8 and human presentation including tumors of the human lung, colon, and anterior chest, such as thoracic carcinoma MCF-7. It has activity to inhibit a wide range of tumors including but not limited to these.

The efficacy of the compositions of the present invention in inhibiting tumors is such that the compositions of formulas ■ to ■ inhibit tumors in host animals. Show that it should also be useful for controlling.

Therefore, the composition of the present invention 1, the method for producing the composition of the present invention, and the method for inhibiting viruses and tumors. Methods for using the compositions of the invention to control viruses and tumors include , destroy the virus and thereby achieve the object of the present invention. Diseases, disorders and symptoms caused by or related to tumors It is effective in controlling pleasure.

(Example) The present invention will be explained by examples. The examples are not intended to limit the scope of the invention. It's not something you do. In conjunction with the detailed general description above, the embodiments further provide that the present invention The following provides a clear understanding and outlines methods for making the compositions of the present invention.

The following examples illustrate compositions, processes of the invention to meet the claimed objects of the invention. and a preferred embodiment of the method. Raw materials and examples in examples that do not show preparation methods and reagents are commercially available from vendors known in the art, such as drug suppliers.

Example l Preparation method of compound (1): 20009” Collected red algae, Desmia hornemannio from Lamb, 5000ml! Add 2000 Fffj of acetone to the container and stir the mixture vigorously. A slurry was made.

The slurry was filtered to obtain an acetone extract. Concentrate the acetone extract under vacuum at room temperature. The mixture was concentrated and mixed with 100 ml of methylene chloride in a separatory funnel. methylene chloride Both fractions were taken out and concentrated to obtain 13.29m of crude oil.

The methylene chloride extract was chromatographed on a silica gel column at 7:3 It was eluted with hexane-acetone.

Compound (X): C.I. (X) The main fraction containing the alcohol derivative of compound (1) shown in Teflon, 1 mz acetic anhydride and o, iml on topography grade silica gel column. ! It was mixed with pyridine to form aceftv. Remove excess acetic anhydride and pyridine. The oil is removed by evaporation under moderate heating and reduced pressure, and this is poured into a 1:1 ratio Xane: Chromatography on silica gel using ethyl acetate elution solvent and purified to give a pure yellow of the formula I got it right.

The exact structure of this compound is still unknown.

Preparation method of compounds (2) and (3): Collected 2000g of red algae Desmia hornemanni and produced a volume of 5000F71/ Add 2000 me of acetone to the container and stir the mixture vigorously. I made slurry.

The slurry is filtered to obtain an acetone extract. Concentrate the acetone extract under vacuum at room temperature. Shrink it in a separatory funnel to 100771! of methylene chloride. methylene chloride Take out the fraction.

Concentration gave 18.29m of crude oil.

The methylene chloride extract was chromatographed on a silica gel column 7:3 It was eluted with hexane-acetone.

Halogen of the formula ■ shown below as in the formula ■ (where X is CI or Br) A volatile oil fraction containing the derivative was obtained.

■ A sample (47,2m9) of compound (Y) of formula B was diluted with io%KOH for 30 minutes at room temperature. - Treated with MeOH, two main spots (CHC13 ) gave a reaction mixture with Separate this mixture with chloroform using a tic Ketal composition f31 (Rf = 0.3, IB, 5m9, 53.4 %), and vinyl ether composition f2) (Rf=o, s, 8.5m9.28 ．． 6%).

Ketal (3) has the following properties: oil content, λmax (EtOH) 235n m(E79oO);IR(:yilm)2960°2830.1660,163 5.1470,1370,1270.1190,1175%1150.1110 ．． 1055.870α iHNMR (CC14) δ7.03 (IH, d, J=1 2H2), 6.65 (IH, dd, J=10.0.5.'2H2), 6.00 (IH, d, J=10.0H2), 5-06 (6H, S'), t, 5o (5H ,S), 1.30(6H,S), Vinyl ether (2) is a two-oil component, λmax(EtOH)2, with the following properties: 35 (E6000), 218 nm (E14000) +IR (Fi lum) 2960.1665.1525. ．． 1580% 1400, 1285.10 80.84 oc! 11-'i HNMR (CC14) δ6.97 (IH, d , J=3.2 H2), 6°53 (I Hldd, J=10°0H2), 6 , DO (1iL d, J = 10 H2), 5.31 (1H,.

d, J=2. oH2), 4.25 (IHld, J = 2.0 H2), 153 ( 3H,S), 1.30 (6H%S), Example 3 Alternatively, the ketal (3) can be obtained by adding 55 mg of a sample (9.3 mg) of compound (Y). Treat with N methanol 1 (OH (1 m/?)) and leave at room temperature for 20 hours. It can be manufactured by After adding 5.0 ml of water, the mixture was diluted with CH Extracted with 2Cl2. Dry the Chzolg solution with sodium sulfate and concentrate. Reduction gave the ketal (31 (6, 4777 g, 87-97%)).

Example 4 Preparation method of compound (4): Ketal t31 (6, s’mg) produced by the method of either Example 2 or 3 ) was dissolved in MeOH of o, imz and 5N HCl of o, imz. 1 After standing for 0 minutes, the solution was extracted with CHCl+ to form an oil with the following properties: 4.5777 g (79.4%) of (4) was obtained: λma, x (Et OH ) 205 (E 7500), 234 nm (E 9200): In (filter) ) 2980% 2940.2875, 1695, 1665, 166°0, 1 600.1400, 1350, 1260, 935.84ot7R; HNM R (CDCL3) 7.50 (IH1d%J=3.oH2), 6.77 (IH , dd, J=10.0%5.0H), 6.12(IH, d%J=10.0%H2 ), 2.50 (3H, S), 1.35 (6H, S).

Example 5 Preparation method (5) of epoxide compound (5): Compound (X) (157,8777g) , 2 mt of l% KOH/J (20, and 2 ml of dioxane mixture was heated under reflux for 2 hours. Dioxane t-[5 reaction mixture after removal with EE was extracted with chloroform. Transfer the extract to silica gel and place it on a column using chloroform. sl, sm as raw material of 62mQ and light yellow oil with the following characteristics. q (46,8%) x Hoxy) - (51ヲ?8*: (d) 31. 2° (CO, 64, MeOH); IR (film) 2975% 2925.287 5.1670.1650.1470, 1420.1255.1240, 1150 , 915.105.880, 820.8001-”; 1HNMRδ6-57 (IH, dd, J=9.0.2.0H2), 6-57 (IH%dd, J=2 ．． 0.0, 4 H2>, 5.92 (IH-d-J=9.0H2), 3.62 ( IH-m)%2.85(IH%ddsJ=6.8,4.3Hz)%2.26(1 H, dd-J=6.82.0H2), 1.25(+H,S)% 1.18(3H ,S).

Example 6 Preparation method of composition (6): Collected 2000g of red algae Desmia and Hornemannio and put them in a 5000mf container. Add 2000mH acetone to the container and stir the mixture vigorously to form a slurry. - created.

The slurry was filtered to obtain an acetone extract. Concentrate the acetone extract in vacuo at room temperature. The mixture was concentrated and mixed with 100 m/ml of methylene chloride in a separatory funnel. methylene chloride The fractions were collected and concentrated to obtain 15.29m of crude oil.

The methylene chloride extract was chromatographed on a silica gel column, 7:3 The elution was carried out with hegisan-acetone. A volatile oil containing composition (6) was obtained.

Examples 7 and 8 Method for preparing compositions (7) and (8): I Composition (6) oil content 0.349m was added with lO% KOH (1mg ) was added slowly without stirring. The mixture was left at room temperature for 30 minutes. blend was chromatographed on a silica gel column using 15:2 hexane/acetic acid. A mixture containing 33.8 m9 of (7) and 190 mg of (8) I got it. Furthermore, 110F? This mixture of i+lF was applied to a Ropar 5t-60 column. Above, 34.3 mg of (8) was separated using 6:1 hexane/ethyl acetate. Obtained.

Composition (7) was a pale yellow oil: IR (film) 3260.3060 ．． 2975.2940.2875.2110.1665.1630.1475. 1400.1380.1360.1305.1255.1180.1135゜1 085.985, 950.930, 910 and 84001″″10 Composition (8) was a pale yellow oil: IR (film), 3050.2970 ．． 2940.2870.1670.1640.1480.1400,1380. 1360.1250.1200, 1170.1130% 1020.960,9 10% 840α.

(Antiviral and antitumor activity of the compound of the present invention) Antiviral power of compound (1) 1) Remove the 75d culture flask from the incubator. .

(These were plated the previous week) 2) Aspirate the medium.

3) Wash with lQmz of PBSA or bag of saline water.

4) Aspirate.

5) Wash once again with 10ml of PBSA or saline water.

6) Aspirate.

7) 2ml! of trypsin/EDTA. Culture until cells separate ( 10-20 minutes).

8) Once separated, shake vigorously, immediately add 8 mg of medium, and shake again. do.

9) Count the number of cells (add 0.IF?+1! to the cell suspension of 0.5 F77/ Add Rivanpuru. Wait 5-10 minutes. With a hemocytometer, measure 4 corners and 5 x 5 rows. Measure the center section. Blue color indicates dead cells. 10mg total in culture flask Cells = number of viable cells x 6 x 105) 10) Add 8 x 10’ cells to another 75 d culture flask to prepare cell liner. hold the button. Bring to 30'WIl with fresh medium.

11) Add IX 10' to each well (6 silanyl per culture dish) for assay. Add cells. Make each well 2 mg with fresh medium.

12) Culture at 37°C overnight.

Day 2: 1) Aspirate the medium.

2) Add 0.5 ml of medium containing 200 pfu of H8V-I.

3) Culture for 102 hours with slight shaking.

4) Add 2 mg of MC-4000 to each well.

5) Place the dinuk on the surface and press it down. (1 mg/day for crude extract) disk, 0.5mQ/disk, 0゜25mQ/Dinuku, etc.) 6) Culture for 48 hours. (Make sure the plate is level) Day 4: 1) Add 2 mg of neutral red medium.

2) - Cultivate overnight.

Day 5: l) Read the wells. For example, in the case of compound 16 (++) of 2I, 16 is the diameter Represents the zone of cytotoxicity in mm (6 mm to 16777777), ( 10) indicates the degree of inhibition of bed formation, complete inhibition (10++), and slight inhibition around the outside of the well. Bed formation (++), constant inhibition (+), doubtful inhibition (+/-), no inhibition -) , and not indicated for complete cytotoxicity.

Start the cycle again on day 1.

1 prescription culture medium 1/ Gipko Mem (plus non-essential a, a and are salt) 2.2q, NaHCO3 5 oml calf serum 105 units penicillin 50mg streptomycin MC-4000 500777/V/W4000Cp8 Methyl cellulose neutral red Culture medium (Always make fresh products) 500777/ Half volume culture medium (double concentration) 500mg 4% V/W15 (!I)8 Methylcellulose 100777g Neutral Red De (from stock solution) back G salt solution ll! Add the following to distilled water: 8.009 NaC1 0.40gKCI O, 159KHz PO4 0.29gNa2 HPO4・7H202.0ml 1% phenol red i, iog glucose 2g Difco 1-25B5A in 100ml bag of saline water l1! Add the following to distilled water: 8.009 NaC1 0.20g KCl 1.50V NazHPO4・7H200, 25gKHzPO4 autoclave Day 5 results for composition (1) show that there is a constant antiviral effect for this compound. It shows an activity of 16 (++). Compound (1) can be used in plaques of 25 to 80 virus cells. Minimum effective amount of antiviral activity of 50 to 100 micrograms per forming unit shows.

The following assay method was used to demonstrate the antitumor potency of compounds 1-7.

L1210 and P388 mouse leukemia cells; ileocecal agamma denocarcinoma H CT-8i and lung carunno-7A 549;+[jJ toxicity assay, 2 4-well plate screening assay and tube assay protocol Le.

Materials used Media - such as glucose and pyruvate (Biologos, Incoholv-John) Rahi 10 Chiuma Serum (Biologos Inc.), 1:1.0 Dulbecco's containing μQ/ml', r':/tamycin (Gipco) ◇ Cells-Ll 210 and p-388 mouse 7 leukemia cells (American type Calcium) Char collection) Add to the medium at a concentration of 5 x 10% cell number/mz. What I got. Sterilized 24-well culture plate for screening (Nunk) or 12X75777777 glass culture tube for tube assay (Becton Dickinson). Microdispenser with 1-5 μl increments (dosage) Lamond, Scientific, Company (Pennsylvania)) Lumo ).

Fin pipette with 5 to 50 μl increments and 50 to 200 μ7? A sample of the composition to be subjected to the FinBi1 assay with increments of in amounts of 200 py/mt and 1.00 pt/mt to perform screening. Add to each well or tube. For the DDC of known active compounds, 100 pq 7/, ml or more for G-Finding assay Using a logarithmic concentration of 0.01/J g/ml; when measuring the range, the maximum Five concentrations are used between the highest and lowest active concentrations.

2. In each well or tube, add 5X10P' cells N in medium, suspension 2. Om 7? Add. Cover the tube loosely with parafilm.

Incubate for 48 hours in a 35-inch CO□ incubator.

4 Visually read the plate using an inverted microscope while comparing with the solvent control group. . Determine the activity as follows: 0 = 90-100 chis vs. control growth 1+ = vs. 75-89 cm for the growth of light 2+ = 50-74 relative to control growth 3+ = 25-49% of control growth 4+ = 25 chi against control growth Repeat for all positive samples using the tube assay method.

5 For tube assay, mix one tube well by swirling, o, sm Take out the diluted solution of n and f in 9.5 ml of accuvette (Kurtil). (isoton-curutyl), while being careful not to create excessive bubbles. Just before counting, invert and mix thoroughly. Count on a Kurtil counter (set the counter to 0. Set to count the solution of '5Fnl); Therefore, the total number of cells/ml in the original assay tube is converted.

Positive control - pincristin or pincristin in aqueous solution Final concentration (2 μl/incubation) of Hygin°Pastein or Pinkrintin control group. ).

Solution concentration Added amount Final concentration of test 10m9/ml 2711 10p9/me1mg/mE2/At 1p9/ ME0.177797ml 2 pl O, 1/J g/meO, [l 1 mQ/ml 2 pi 0.001 pQ/ml Note: For solvents outside waterfowl, allow the solvent to evaporate from the tubes or wells in a hood. and.

Chloroform and butanol are not suitable for plastic 24-well plates. No need to use a glass tube.

for the last two wells of each plate or each rack of 72 or fewer tubes. Be sure to set up two control groups of solvent in four wooden tubes. On the other hand, a series of plates or 4 wells or tubes containing only medium and cells will be established. When using more than 200 μl of aqueous solution, dry the sample and Adjust the density of the background to the desired density.

Results of the above assay; Formula 11. ．． ■, ■ and) 1 compound is P-888 mouse Cytotoxic to leukemia cells in vitro, 57.70.70 and The appearance of 97ml of 5p indicates the above ID5o (+f411 cell count 050 lethal dose) . Compound ■, V%11 and t114L-1210 murine leukemia cells Total cytotoxicity in vitro, 15% each at 200 micrograms/ml The survival rates were 17%, 41% and 25%. Compound 11 is T, -121 The apparent value of 0.7 micrograms per milliliter for 0 murine leukemia cells is above I. Indicates D5o. The table below summarizes these results.

table Composition -2"-1" Cell JIDso Nifumuraru) p-388A349 HCT- 8 15pt/m! 20p LSI/ml 1071pml! 2　5pQ/ml　 50p9/ml 20p9/m15 10pQ/ml 30pQ/ml 30p Q/m14 5 p 9 / 7771 5 / A 9 / ml 5 p Q /77715 10p9/ml 1p9/ml 5p (1pm!60-5 p 9 / ml 5 A9 / F771 5 μQ 7m 171 p 9 /F771 3 p g/7771 0,3 /j 9 / 777 /8 5 P9/ml　5p9/ml　5p9/ml　5p9/ml　5p9/ml　5p9/ml　5p9/ml　5p9/mL　5p9/ml Without being limited to the examples and suggested applications, modifications may be made without departing from the spirit of the invention. can be changed. Example 1, such as fluorinated cyclohexadienone Other derivatives of cyclohexadienones are similar to the preferred embodiments described above. It may be noted that it has antiviral or antitumor activity. moreover.

The compositions described herein may have other useful uses, such as for pain relief. stomach. That is, if the invention falls within the scope of the appended claims and the like, This covers modifications and variations of the present invention.

Submission of Translation of Written Amendment (Article 184-7, Paragraph 1 of the Patent Law) Clause 1986, April 18th

Claims (1)

[Claims] 1 General formula: ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ composition of (If R1 groups are the same or different, hydrogen or lower alkyl group, X is lower alkyl group, alkyl, hydroxy, lower alkoxy, lower acyloxy, fluor, chlor, Bromine or iodo group, and R2 is hydroxy, lower acyloxy group, halogen (lower acyloxy groups or halogens). Composition according to claim 1, characterized in that 2R1 is a methyl group. Claim 1, characterized in that 3X is C1 and R2 is an acyloxy group. Composition according to item 1. Claims characterized in that 4X is methyl, methoxy, chlor or bromo Composition according to paragraph 1. Type 5: ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ Composition of. 6. As an active ingredient, an antiviral effective amount or amount of one or more of the compositions according to claim 1. Antiviral compositions consisting of a non-toxic filler or diluent and a pharmaceutically approved non-toxic filler or diluent. 7. As an active ingredient, an antiviral effective amount or amount of one or more of the compositions of claim 3. Antiviral compositions consisting of a non-toxic filler or diluent and a pharmaceutically approved non-toxic filler or diluent. 8. As an active ingredient, one or more antiviral effective amounts of the composition of claim 5. Antiviral compositions consisting of a non-toxic filler or diluent and a pharmaceutically approved non-toxic filler or diluent. 9 viruses with an antiviral effective amount of one or more of the compositions of claim 1. A method of inhibiting a virus in a host comprising causing 10 viruses in contact with an antiviral effective amount of one or more of the compositions of claim 3. A method of inhibiting a virus in a host, which consists of exposing the host to a virus. 11 viruses with an antiviral effective amount of the composition of claim 5. A virus inhibition method consisting of. 12 As active ingredients, an antitumor effective amount of one or more of the compositions of claim 1 and An anti-tumor composition comprising a non-toxic bulking agent or diluent approved by the Pharmacy and Pharmacy Act. 13. Contacting the tumor with an antitumor effective amount of one or more of the compositions of claim 1. A method for inhibiting tumors in a host comprising: 14. Contact red algae with a suitable non-polar organic solvent to obtain an extract using the solvent. characterized in that it consists of each step of isolating the compound according to claim 1 from the output material. A method for producing a compound according to claim 1. 15 Red algae is brought into contact with an appropriate first organic solvent to obtain an extract thereof, and an appropriate second organic solvent is obtained. Mix with organic solvent and extract further, formula Z: ▲ Contains mathematical formulas, chemical formulas, tables, etc. ▼ Solvent extract containing alcohol derivatives of (Z) and isolate the alcohol derivative of the compound of formula Z by chromatography, Each step of acetylating the alcohol derivative and isolating the compound of formula Z A method for producing a compound according to claim 5, comprising: Claim 15, characterized in that the red algae is Desmia hornemannii. method. 17 The first solvent is acetone, the second solvent is methylene chloride, and the compound of formula Z alcohol derivatives in a mixture of hexane and acetone on a silica gel column. Isolate by chromatography by elution and silica gel column chromatography The main areas from the graph were further thinned using flash chromatography. Separated on a layer chromatography grade silica gel column, derivatives of compounds of formula Z and acetylation of the alcohol derivative with a mixture of acetic anhydride and pyridine. 16. A method according to claim 15, characterized in that: 18 general formula: ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ composition by, However, R is ▲There are mathematical formulas, chemical formulas, tables, etc.▼;▲There are mathematical formulas, chemical formulas, tables, etc.▼;▲Numbers There are formulas, chemical formulas, tables, etc. ▼; ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼; ▲ Mathematical formulas, There are chemical formulas, tables, etc.▼;▲There are mathematical formulas, chemical formulas, tables, etc.▼;▲Mathematical formulas, chemistry There are formulas, tables, etc. ▼; ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼; or ▲ Mathematical formulas, chemical formulas, etc. There are scientific formulas, tables, etc.▼Here, X is O, S or NHR1; X22 is hydrogen, halo Gen, lower alkyl group or alkylsulfenyl group; Y is O or S; Z is O , S or NR1; both R1 groups are the same or different and hydrogen or lower alkyl is a group. R2 is H or a lower alkyl group; and n is 1 to 5. 19R is ▲There are mathematical formulas, chemical formulas, tables, etc.▼;▲There are mathematical formulas, chemical formulas, tables, etc.▼;▲Numbers There are formulas, chemical formulas, tables, etc. ▼; ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼; ▲ Mathematical formulas, It consists of chemical formulas, tables, etc.▼; and ▲mathematical formulas, chemical formulas, tables, etc.▼ choose from the group Composition according to claim 18, characterized in that: 20R is ▲There are mathematical formulas, chemical formulas, tables, etc.▼ or ▲There are mathematical formulas, chemical formulas, tables, etc.▼ Composition according to claim 18, characterized in that: 21R is ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ or ▲ There are mathematical formulas, chemical formulas, tables, etc. A composition according to claim 18, characterized in that it is ▼. Claims characterized in that 22R is ▲There are mathematical formulas, chemical formulas, tables, etc.▼ Composition according to paragraph 18. Claims characterized in that 23R is ▲There are mathematical formulas, chemical formulas, tables, etc.▼ Composition according to paragraph 18. 24. As an active ingredient, one or more antitumor effective amounts of the composition of claim 18. and a non-toxic filler or diluent approved by the Pharmacy Act. 25 As an active ingredient, an antitumor effective amount of one or more of the compositions of claim 19 and a non-toxic filler or diluent approved by the Pharmacy Act. 26. Contacting the tumor with an antitumor effective amount of one or more of the compositions of claim 18. A method for inhibiting tumors in a host comprising: 27. Contacting the tumor with an antitumor effective amount of one or more of the compositions of claim 19. A method for inhibiting tumors in a mammalian host comprising: 28 Collect red algae, contact this red algae with an appropriate organic solvent, and extract with the solvent. and synthesize the compound according to claim 16 from the extract. A method for producing a compound according to claim 16, comprising the steps of separating. 29. Contacting the tumor cells with an antitumor effective amount of the composition according to claim 1. To treat cancer cachexia caused by the presence of a tumor in the host consisting of treatment for 30 The tumor cells are contacted with an anti-tumor effective amount of the composition according to claim 18. Treatment of cancerous cachexia caused by the presence of tumors in the room consisting of treatment for. 31. Contacting the tumor cells with an anti-tumor effective amount of the composition according to claim 19. Treats cancer cachexia caused by the presence of a tumor in the host consisting of treatment for.