JP3507511B2 - Pharmaceutical compositions of arglabin and arglabin derivatives - Google Patents

Description

BACKGROUND OF THE INVENTION Cancer is the leading cause of death in the United States and affects people around the world. Surgery, radiation therapy, and chemotherapy are the most widely used treatment modalities. Chemotherapeutic agents create conditions in cells that limit cell growth and replication. DNA synthesis can be inhibited by purine synthesis, pyrimidine synthesis, conversion of ribonucleotides to deoxyribonucleotides, antimetabolites, intercalation, or cross-linking. RNA synthesis may be inhibited by, for example, antagonists.
Protein synthesis can be inhibited, for example, by agents that deaminate asparagine. Furthermore, agents that inhibit the function of microtubules can be used as chemotherapeutic agents.

Chemotherapeutic agents also typically affect neoplasms and rapidly growing cells of normal tissues such as bone marrow, hair follicles, and intestinal epithelium. Anorexia, nausea, vomiting, diarrhea, suppression of bone marrow function, and hair loss are usually some of the negative effects associated with chemotherapy. The benefits of developing chemotherapeutic agents that are effective anti-tumor agents with minimal toxicity are great.

SUMMARY OF THE INVENTION It has been found that algrabine, and various algrabine derivatives, can function as effective chemotherapeutic agents with fewer side effects than are commonly brought about by the use of other chemotherapeutic agents.

In one aspect, the invention features a pharmaceutical composition in unit dosage form suitable for treating cancer in humans. The composition consists essentially of about 40 mg to about 480 mg of algrabin or a derivative thereof. A unit dose of this composition is, for example, from about 175 mg to about 315 mg, or about
It can be 240 mg to 280 mg arglabin or a derivative thereof. Algrabin or a derivative thereof can be used in the manufacture of a medicament for treating cancer. The composition is
It can be used in the manufacture of a medicament for treating cancer. The composition is useful for treating a wide variety of cancers including, for example, breast cancer, colon cancer, rectal cancer, gastric cancer, pancreatic cancer, lung cancer, liver cancer, ovarian cancer, pancreatic cancer, esophageal cancer, leukemia, and lymphoma. is there. This composition is particularly useful for treating lung cancer, liver cancer, and ovarian cancer.

Dimethylaminoarglabin, or a pharmaceutically acceptable salt thereof, is a particularly useful algrabin derivative that can be used as a pharmaceutical composition. Dimethylaminoarglabin or a pharmaceutically acceptable salt thereof can be lyophilized.

The invention also features a composition that includes a first chemotherapeutic agent comprising algrabin or a derivative thereof and a second chemotherapeutic agent. The second chemotherapeutic agent is not arglabin or its derivative. This composition is effective in inhibiting tumor growth in humans. A particularly useful algrabine derivative is dimethylamino algrabine or a pharmaceutically acceptable salt thereof. The second chemotherapeutic agent is, for example, cyclophosphamide, sarcolys (sarcolys).
in) or an alkylating agent such as methylnitrosourea, an antimetabolite such as methotrexate or fluorouracil, a vinca alkaloid such as vinblastine or vincristine, an antibiotic such as rubidomycin, or a platinum complex such as cisplatin. Two or more chemotherapeutic agents can be included in the composition along with algravin or a derivative thereof. This composition can be used in the manufacture of a medicament for treating cancer.

The invention also features compounds that inhibit tumor growth in mammals. These compounds are selected from the group represented by formulas I, II, III, IV, V, and VI: In the formula, RR ₁ is NHCH ₂ Ph or N (CH ₂ CH ₂ ) ₂ O, and RR
₂ is NHCH ₂ Ph, N (CH ₂ CH ₂ ) ₂ O, N (CH ₃ ) ₂ or a pharmaceutically acceptable salt thereof, and X is OH or Cl.
Is. These algrabine derivatives include dimethylamino epoxy algrabine, dibromo algrabine, algrabine chlorohydrin, 11,13 dihydro algrabine, benzylamino algrabine, morpholine-amino algrabine, benzylamino epoxy algrabine,
Morpholine-amino epoxy algrabine, epoxy algrabine chlorohydrin, or pharmaceutically acceptable salts thereof are included. Formulas I, II, III, IV,
The compound of V or VI can be used in the manufacture of a medicament for suppressing tumor growth in a mammal.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present description, including definitions, will control. The materials, methods, and examples are
It is for illustration only and has no limiting meaning.

Other features and advantages of the invention will be apparent from the following detailed description, and from the claims.

BRIEF DESCRIPTION OF THE FIGURES FIG. 1 illustrates the synthesis of arglabin derivatives 2-9.

FIG. 2 illustrates the synthesis of arglabin derivatives 10 to 13.

FIG. 3 illustrates the synthesis of arglabin derivatives 14a-14d, 15a-15d, and 16.

FIG. 4 illustrates the structures of compounds 17-21.

FIG. 5 shows the effect on the survival rate of the transformed cells when the concentration of dimethylaminoarglabin hydrochloride was increased.

FIG. 6 shows the effect of increasing the concentration of dimethylaminoarglabin hydrochloride on the growth of transformed cells.

FIG. 7 shows the effect of increasing the concentration of dimethylaminoarglabin hydrochloride on the viability of normal cells.

FIG. 8 is a plot of the spectral index of naphthol degradation products. Figure 8A shows no drug, Figure 8B
Is when the drug is put.

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention provides novel compounds that suppress tumor growth in humans. These compounds can be synthesized from the parent compound algrabine (Figure 1), which is isolated from Artemisia glabella. A variety of arglabin derivatives can be created using a range of chemistries. For example, epoxy arglabin can be produced by epoxidizing a trivalent substituted olefinic double bond with peracetic acid. Dichlorohydrin can be prepared by treating epoxyarglabin with an ether-acetone hydrochloric acid solution. Dibromoarglabin can be produced by reacting algrabin with Br ₂ and carbon tetrachloride. Algravin chlorohydrin can be produced from algravin by reacting with a methanolic hydrochloric acid solution. Epoxidation of arglabin chlorohydrin with peracetic acid and chloroform gives the epoxy arglabin chlorohydrin which can be separated by chromatography. Algrabindiol, its isomers,
The diene type and the diene type can be produced by hydrolyzing algravin. Epilagrabine, the 1,10 epimer of arglabin, can be prepared by treating arglabin diol with POCl ₃ . Benzylaminoarglabin and benzylaminoepoxyarglabin can be produced by treating algrabin and epoxyarglabin with benzenamine. Dimethylaminoarglabin and dimethylaminoepoxyarglabin can be produced by treating algrabin and epoxyarglabin with dimethylamine. Morpholine-aminoarglabin and morpholine-aminoepoxyalgrabin can be produced by aminating algorabin with morpholine. Pharmaceutically acceptable salts of these compounds can be produced according to a conventional method and can be used as an antitumor agent. For example, dimethylaminoarglabin hydrochloride and dimethylaminoepoxy algrabine hydrochloride are
It can be produced by hydrochloric acid. Dihydroarglabin can be produced by treating algrabin with ethanol and H ₂ / Ni. The various arglabin derivatives described above are shown in Figures 1-4.

The invention also relates to a method of suppressing tumor growth in a human patient diagnosed with cancer, comprising administering to the patient arglabin or a derivative thereof. This method
Generally, it can be used to treat cancers such as breast cancer, colon cancer, rectal cancer, gastric cancer, pancreatic cancer, lung cancer, liver cancer, ovarian cancer, pancreatic cancer, esophageal cancer, leukemia, and lymphoma, but lung cancer, Certain types of cancer, such as liver cancer and ovarian cancer, are particularly amenable to this treatment. The compounds are topically, orally, intravenously, intraperitoneally, intrapleurally, intrathecally, depending on the type of cancer and various indicators of the patient.
It can be administered subcutaneously, intramuscularly, intranasally, by inhalation, or by suppository. For example, intraperitoneal administration may be used for patients with ascites. Intrapleural administration can be used for certain lung cancer patients. The suppository can be used for patients with rectal cancer. Algrabin or its derivatives are
In a daily dose, about 40 mg to about 480 mg, preferably about 175
mg to about 315 mg, more preferably about 240 mg to 280 mg can be administered. A typical dose range is about 0.5 m
It is from g / kg to about 7 mg / kg. About 20 for extremely dangerous symptoms
Up to mg / kg arglabin or its derivatives can be administered. When administered, these compounds can act as anti-tumor agents to inhibit tumor growth or cause tumor regression.

Without being constrained by specific biochemical mechanisms, these compounds can eliminate cancer cells or inhibit cancer cell growth by interfering with farnesylation of proteins such as the ras protein. ra
The s gene is a protooncogene involved in many types of human cancers such as colorectal cancer, exocrine pancreatic cancer, and bone marrow leukemia (Barbacid, 1987, Ann. Rev. Biochem. 56: 779). Approximately 20% to 30% of all human tumors may be due to activation of the ras protooncogene. The ras gene is ras
It constitutes a multigene family that transforms cells by the action of a 21 kDa protein called p21 (also referred to herein as "ras"). ras is G
It functions as a G regulatory protein that hydrolyzes TP to GDP. When inactivated, ras binds to GDP. When the growth factor receptor is activated, ras exchanges with GDP and changes its conformation. With GTP bound, wild-type ras binds the activated growth factor receptor signal to downstream mitogen effectors. The intrinsic GTPase activity of ras causes the protein to eventually return to its inactive GDP-bound state.
In tumor cells, mutations in the ras gene
The regulatory function is lost and the growth stimulatory signal is constantly transmitted, resulting in activation of oncogenes.

To function normally and as an oncogene, ras must localize to the plasma membrane by a process that depends on proper post-translational processing of ras (Hancock, 1989, Cell 57). : 1167). In the first step in the post-translational processing of ras, the farnesyl group is attached to the cysteine residue at position 186 of this protein by reaction with farnesyl pyrophosphate. The action of a specific protease then cleaves the three amino acids at the carboxyl terminus of this protein. In addition, the carboxylic acid end is converted to a methyl ester by alkylation with a methyl group.

Post-translational modifications of ras are often mediated by an amino acid sequence motif called the "CAAX box." In this sequence motif, C represents cysteine, A represents an aliphatic amino acid, and X is another amino acid such as methionine, serine, or glutamine. Depending on the specific sequence of the CAAX box, this motif serves as a signal sequence for farnesyl protein transferase, or geranylgeranyl protein transferase, which catalyzes the alkylation of cysteine residues in the CAAX box sequence. Farnesylation of ras requires proteolytic processing, palmitoylation, and strong binding of the ras protein to the cell membrane.

Without farnesylation, the oncogenotype of ras cannot transform cells as oncogenes. Indeed, inhibitors of farnesyl protein transferase have been shown to block the growth of ras-transformed cells in soft agar. Therefore, farnesyl protein transferase inhibitors, and inhibitors of general ras activity, are believed to be useful as anti-cancer therapeutic agents for many types of cancer (Gibbs et al., 1984, Proc. Natl. Acad. Sc.
i. USA 81: 5704-5708; Jung et al., 1994, Mol. Cell. Biol. 14:
3707-3718; Predergast et al., 1994, Mol. Cell. Biol. 14: 419.
3-4202; Vogt et al., 1995, J. Biol. Chem. 270: 660-664; and Maron et al., 1995, J. Biol. Chem. 270: 22263-22270).

As described below, arglabin and its derivatives are
It is thought to inhibit farnesylation of proteins.

In another embodiment, a pharmaceutical composition comprising about 40 mg to about 480 mg, preferably about 175 mg to about 315 mg, more preferably about 240 mg to about 280 mg of algrabin or a derivative thereof is provided in the form of a unit dose. It The dosage is
The daily dose may be divided into 2 to 4 doses. A typical dosage of these pharmaceutical compositions is from about 0.5 mg / kg to about 7 mg / kg. In special circumstances, up to about 20 mg / kg can be administered. Lyophilized dimethylaminoarglabine and pharmaceutically acceptable lyophilized salts such as dimethylaminoarglabine hydrochloride are particularly useful as pharmaceutical compositions. The optimal concentration of algrabine or a derivative thereof in a pharmaceutically acceptable composition will depend on many factors, including the preferred dose of the compound to be administered, the chemical properties of the compound used, the formulation of the compound excipient, and the route of administration. Can vary depending on the elements of. The optimal dosage of the pharmaceutical composition administered will also depend on variables such as the type and extent of cancer metastasis, the general health of the particular patient, and the relative biological effectiveness of the selected compound. To do. These compositions can be used, inter alia, for the treatment of lung, liver and ovarian cancers, although other cancers such as breast, rectal, colon, gastric, pancreatic or esophageal cancers can also be used in the composition. Effectively treated by the thing. Furthermore, hematopoietic cell carcinomas such as leukemias and lymphomas can also be effectively treated.

The compounds of the present invention can be formulated into pharmaceutical compositions by mixing with non-toxic pharmaceutically acceptable excipients or carriers. Such compounds and compositions are for parenteral administration, especially in the form of an aqueous solution or suspension in physiological buffer, and for oral administration, especially in the form of tablets or capsules. For intranasal administration, it can be prepared especially in the form of powder, nasal drops, or aerosol. Compositions for use in other routes of administration can be prepared, if desired, using conventional methods.

The compounds of the present invention can be conveniently administered in the form of unit doses and also by methods well known in the art of pharmacy, such as Remington's Pharmaceutical Science ( Mack Pub.
It can be prepared by the method described in Co., Easton, PA. Formulations for parenteral administration may include, as general excipients, sterile water or saline, polyalkylene glycols such as polyethylene glycol, oils and fats of plant origin, hydrogenated naphthalene, and the like. In particular, biocompatible, biodegradable lactide polymers, lactide / glycolide copolymers, or polyoxyethylene-polyoxypropylene copolymers are examples of excipients for controlling the in vivo release of the compounds of the invention. is there. Other suitable systems for parenteral delivery include ethylene-vinyl acetate copolymer particles, osmotic pumps, implantable infusion systems, and liposomes. Formulations for administration by inhalation may include excipients such as lactose, if desired. The formulation for inhalation may be, for example, an aqueous solution containing polyoxyethylene-9-lauryl ether, glycocholic acid, and deoxycholic acid, or an oily solution for administration in the form of nasal drops. The compounds, if desired, can also be formulated as gels for intranasal application. Formulations for parenteral administration also include glycocholate for buccal administration.

The invention also relates to a packaging material and an article of manufacture comprising algravin or a derivative thereof encased within the packaging material. Algrabin or a derivative thereof is effective as a treatment for suppressing tumor growth in humans. The packaging material includes a label or insert that indicates that arglabin or a derivative thereof can be used to inhibit tumor growth in humans. Dimethylaminoalgrabin, and its pharmaceutically acceptable salts are particularly useful algravin derivatives for use as products.

In another aspect, the invention relates to compositions and kits that include a first chemotherapeutic agent that includes algrabin, or a derivative thereof, and a second chemotherapeutic agent. The second chemotherapeutic agent is not arglabin or its derivative. These compositions are effective in suppressing tumor growth in humans. Dimethylaminoarglabin or a pharmaceutically acceptable salt thereof is a particularly useful algrabine derivative. Various types of chemotherapeutic agents can be used in this composition, such as alkylating agents, antimetabolites, vinca alkaloids, antibiotics, or platinum coordination complexes. For example, alkylating agents such as nitrogen mustard cyclophosphamide and sarcosine can be used, although other alkylating agents such as methylnitrosourea are also suitable. Antimetabolites such as the folic acid compound methotrexate or pyrimidine analogs such as fluorouracil or 5-fluorouracil can also be used, as well as vin alkaloids such as vinblastine or vincristine. Antibiotics such as rubidomycin, as well as platinum coordination complexes such as cisplatin, may be suitable chemotherapeutic agents. Many chemotherapeutic agents can be combined with arglabin or its derivatives. For example, vincristine and cyclophosphamide or vincristine and vinblastine can be combined with arglabin or a derivative thereof.

The present invention also provides for administering to a patient a fixed amount of a composition comprising a first chemotherapeutic agent comprising algrabin or a derivative thereof and a second chemotherapeutic agent effective in suppressing tumor growth in humans. , A method of inhibiting tumor growth in human patients. The second chemotherapeutic agent is not arglabin or its derivative. With these compositions, an enhanced antitumor effect can be obtained and the occurrence of metastases can be prevented. In particular, these compositions are useful in defeating drug resistant tumors. These agents may be administered separately or as a mixture. Toxicity can be reduced by administering algravin or a derivative thereof several hours before the administration of the chemotherapeutic agent. These compositions may be administered by any route.

The present invention also provides for immunosuppression of a chemotherapeutic agent in a human patient by administering to the patient an amount of arglabin or a derivative thereof that is effective to enhance the patient's immune system when treating the patient with the chemotherapeutic agent. Regarding methods for reducing the effect. The immune system can be strengthened, for example, by increasing the total number of leukocytes, T-lymphocytes, B-lymphocytes, or immunoglobulins.

The invention is further described in the following examples,
It does not limit the scope of the invention as claimed.

Examples Example 1: Isolation of arglabin-smooth w
ormwood), Artemisia Gravela Kar.et Kir. (Arte
misia glabella Kar.et Kir.) is a perennial plant that widely propagates in the dry steppe plateau of Kazakhstan. The aerial parts of A. glabella, including leaves, buds, florets, and stems, contain sesquiterpene lactones such as algrabin throughout the plant's entire growth stage (Table I).

Sesquiterpene lactones were extracted from dried plants using various solvents (Table II). It was found that higher yields were obtained by extracting the lactone three times with chloroform from the flowering plants at 45-50 ° C.

A backflow type continuous extraction device, a filling device, and an extraction device consisting of three types of containers separated from the external environment were used for extraction. The container containing the solvent is equipped with a filter and a distillation device having an evaporation device and a concentration device, and has a buffering capacity. The container containing the desiccant is a dryer, cyclone,
It consists of a cooling device, a ventilation device, and a heating device. The container containing the cooling water has a breathable salt pan.
This extraction device includes a deodorizing device, a drain tank, and an extract recovery device.

Artemisia gla Kar.et Kir. (Artemisia gla
bella glabella Kar.et Kir.) dried material about 7.7k
g was placed in the extractor and constantly mixed with the solvent while passing through the extraction column. The solvent moves in a direction opposite to that of the dried plant material, gradually becoming saturated with the extracted material. Once saturated, the solvent was first filtered to remove fragments of plant material and then evaporated.
The filtered plant pieces were recycled in the extractor for re-extraction. The evaporated vapor was sent to the concentrator. The pure solvent was recovered from the concentrator and recycled to the extractor. The agglomerated surface in the concentrator was cooled by the outside air blown from the aeration device, and by the water pumped from the salt pan that previously cooled the water. By evaporative cooling in the salt pan with air, the water can be cooled to a temperature well below the temperature of the outside air.

Extracted material purified from the solvent is tar-like. About 7% of plant material (539 grams) during this process
Was recovered.

This tar-like material was further purified by adding 2 volumes (about 1.08 L) of 60 ° C. ethanol with constant stirring to dissolve the tar. Distilled water warmed to about 70 ° C. was added in an alcohol to water ratio of 2: 1. Stir the tar-alcohol-water solution carefully for 30 minutes, then
Let stand at room temperature for 24 hours or until a precipitate is formed.
The hydroalcoholic solution was filtered through a ceramic filter under vacuum. This procedure was repeated with the precipitate remaining after filtration.

The filtrate was rotary evaporated and the alcohol was vacuum distilled in the form of an azeotrope with water containing 68-70% alcohol. After distilling the alcohol, about 286 grams of purified tar was obtained from this aqueous solution.

The purified tar was applied to a KCK silica gel column using benzene as an eluent and separated into each component by pressure. The benzene fraction was collected and analyzed for algrabin using thin layer chromatography (TLC) (silufol, benzene-ethanol, 9: 1). Fractions containing arglabin were distilled to remove benzene. Algravin at this stage is yellow in color. With a yield of about 11.7%,
33 g of arglabin was produced.

Algravin was recrystallized by dissolving in hexane and heating so that the ratio of product to hexane (w / v) was 1:10. After dissolving arglabin in the solution, the product was vacuum filtered. At the room temperature, the filtrate was separated from the crystals of algrabine. In this step, about 21 g of arglabin was recovered. Algravin is 1 (R), 10 (S)
-Epoxy-5 (S), 6 (S), 7 (S) -Guia-3
It has a structure of (4), 11 (13) -diene-6,12-olide. The stereochemical structure of arglabin was determined by X-ray analysis.

It is the transoid that connects the pentene and heptane rings with the heptane and γ-lactone rings into crystallographically distinct algrabine molecules. Twist angle, respectively, the _{O 3 C 1 C 5 H 5} , and -142 (1) -136
(2) °, and for H ₆ C ₆ C ₇ H ₇ , -167 (2) and -159
(3) °. The pentene ring accepts the 1α-envelope type conformation (ΔC ¹ _S = 2.9 and 1.5 °), and the heptane ring contains the 7α, 1,10β chain (ΔC ⁷ _S = 2.7 and
4.7 °). The methyl group next to the C-10 atom is in the equatorial α-direction. The conformation of the γ = lactone ring was intermediate between the 7α-envelope type and the 6β, 7α-half chair type, but closer to the latter (Δ
C ¹² ₂ = 2.0 and 6.1 °).

The NMR spectrum of arglabin was measured by Varian HA- in CDC1.
Recorded on a 100D instrument. Chemical shifts set the δ value to 0 from the received signal TMC. At 1.34 ppm (epoxy methyl group) and 1.94 ppm (double bond methyl group),
There are two 3-proton singlets. A single proton doublet was recorded at 2.95 ppm (proton in C position) with J = 10 Hz. The triplet of one proton is J = 10
Detected with a center of 3/97 ppm (lactone protons) with Hz. Two 1-proton doublets were obtained at 5.42 ppm with J = 3 Hz and 6.1 ppm with J = 3 Hz (Lactone ring exomethylene), and 1 proton signal at 5.56 (vinyl proton). ) Was obtained. Isolated compounds and related sesquiterpene lactones Arborescien and ludartine
The structure of algrabine (Fig. 1) was confirmed based on the NMR spectrum of.

Overview of the characteristics of arglabin: colorless, melting point approx. 100-102 ° C
(Hexane); [α] ²⁰ _D + 45.6 ° (c 0.3, CHCl ₃ ); IR
Band (KBr) 1760, 1660, 1150, 1125 cm ^-1 ; ¹ H-NMR (4
00MHz, CDCl ₃ ) δ1.34 (3H, s, H-14), 1.94 (3H,
s, H-15), 2.95 (1H, d, J 10Hz, H-5), 3.97
(1H, t, J 10Hz, H-6), 5.56 (1H, br, s, H-
3), 5.42 (1H, d, J 3Hz, H-13a), 6.10 (1H,
d, J 3Hz, H-13b).

Example 2: Algravin Derivatives-To aid readers of this specification, a number is appended to the various compounds below to facilitate identification with the compounds shown in the figures. I am doing it.

Algravin was derivatized with reagents that act on the epoxide and olefin groups of algrabin. Epoxidation of the trisubstituted olefinic double bond of algravin 1 with peracetic acid (Fig. 1) proceeded with high yield and 95% stereoselectivity, and proceeded with 3β, 4β-epoxy algravin 2 (1).
The (10), 3 (4) -diepoxy-guay-11 (13) -ene-6,12-olide is formed. Epoxyargrabine 2 was recovered in a yield of about 65% using silica gel column chromatography with ethyl ether. IR and
Epoxyargrabine 2 using NMR spectrum
The structure of was confirmed.

Overview of the characteristics of Epoxy Algravin 2: melting point 149-151
℃ (Et ₂ O-CH ₂ Cl ₂ ); [α] ²² _D + 94.0 ° (c 1.7, CHC
l ₃ ); IR band (KBr) 1760, 1670 cm ^-1 ; ¹ H-NMR (400 MH
z, py−d ₅ ) δ1.30 (3H, s, H-14), 1.68 (3H,
s, H-15), 3.31 (1H, s, H-3), 4.11 (1H,
t, J 10Hz, H-6), 5.43 (1H, d, J 3Hz, H-13
a), 6.16 (1H, d, J 3Hz, H-13b).

Treatment of epoxyarglabin 2 with an ether-acetone hydrochloric acid solution produces dichlorohydrins 3 and 4 (FIG. 1). It was observed that cleavage of both epoxy groups was obtained with 60% and 95% regioselectivity. Dichlorohydrin 3 and 4 were diluted with water, washed with NaHCO ₃ and Et ₂ O-
Purified by silica gel chromatography using iPr ₂ O in a 1: 1 ratio. The manufacture was determined using IR and NMR spectral data.

Summary of characteristics of dichlorohydrin 3 and 4: 3 (60%)
Melting point 190-191 ° C; [α] ²⁰ _D- 90.1 ° (c 0.34 acetone); IR band (KBr) 3465, 1750, 1670 cm ^-1 ; ¹ H-NMR
(400MHz, py-d ₅ ) δ 1.61 (3H, s, H-14), 1.62
(3H, s, H-15), 4.42 (1H, dd, J10, 7Hz, H-
3), 4.48 (1H, t, J 10Hz, H-6), 5.52 (1H,
d, J 3Hz, H-13a), 6.04 (1H, d, J 3Hz, H-13
b). 4 (5%) mp _{176~178 ℃ (CH 2 Cl 2 -Et} 2 O);
[Α] ²⁴ _D + 23.25 ° (c 0.43, CHCl ₃ ); IR band (KBr)
3680, 1770, 1670 cm ^-1 ; ¹ H-NMR (400 MHz, py-d ₅ ) δ 1.
41 (3H, s, H-14), 1.57 (3H, s, H-15), 3.39
(1H, d, J 10Hz, H-5), 3.42 (1H, s, 10-O
H), 4.05 (1H, d, J 5.5Hz, H-3), 4.40 (1H,
s, 4-OH), 4.55 (1H, t, J 10Hz, H-6), 5.54
(1H, d, J 3Hz, H-13a), 6.21 (1H, d, J 3Hz,
H-13b).

Bromination and interaction with N-bromosuccinimide in aqueous acetone produced another derivative, forming a bromohydrin flexible on the trisubstituted double bond and partial bromination of the exomethylene group. Happened. Treatment of arglabin 1 with Br ₂ and carbon tetrachloride at 0 ° C. yielded 3,4 dibromoarglabin 5.

When arglabin 1 was treated with a methanolic hydrochloric acid solution, a mixture of chlorohydrin 6/7, which was chromatographically separable, was obtained in a high yield in a ratio of about 6: 1 (FIG. 1). At the same time, a partial attachment of the hydrochloric acid component to the exomethylene double bond was observed by the Michael type reaction. Regioisome with high abundance
Epoxidation of r) 6 with peracetic acid in chloroform resulted in a 1: 1 ratio of a mixture of epoxyarglabin chlorohydrin isomers 8/9 which could be separated by chromatography. The structures of unknown chlorohydrins 6 to 9 were confirmed based on the component analysis and the spectrum analysis while considering the result of the epoxide 8 by X-ray.

Approximately 550 mg of arglabin 1 was refluxed with approximately 15 ml of acetonitrile and 1 drop of HBF ₄ for 1.5 hours to produce diol 10 as a major product, which produced isomer 11 and diene 12 in low yield (Fig. 2). ). The reaction was neutralized, diluted with water and extracted with chloroform before column chromatography (10, petroleum-ethyl ether 2: 1; 11, petroleum ethyl ether 1: 1; 12, petroleum ethyl ether 1: 3. ).

Summary of characteristics of diol 10, isomer 11, and diene 12: 10 melting point 184-185 ° C (ethyl ether); [α] ²¹ _D +
72.3 ° (c 0.3 CHCl ₃ ); IR band (KBr) 3440, 1770, 1
680 cm ^-1 ; ¹ H-NMR (400 MHz, py-d ₅ ) δ 1.30 (3 H, s,
H-14), 1.92 (3H, brs, H-15), 4.18 (1H, dd,
J10, 1Hz, H-6), 5.43 (1H, brs, H-3), 5.38
(1H, d, J 3.5Hz, H-13a), 6.14 (1H, d, J 3.5H
z, H-13b). 11 melting point 149-151 ° C (CHCl ₃ -Et ₂ O);
[Α] ²⁵ _D + 108.6 ° (c 0.3 CHCl ₃ ); IR band (KBr) 3
460, 1770, 1670 cm ^-1 ; ¹ H-NMR (400 MHz, py-d ₅ ) δ1.3
5 (3H, s, H-14), 1.92 (3H, brs, H-15), 3.1
0 (1H, d, J 10Hz, H-5), 4.39 (1H, t, J 10H
z, H-6), 5.48 (1H, brs, H-3), 5.44 (1H,
d, J 3.5Hz, H-13a), 6.14 (1H, d, J 3.5Hz, H
-13b). 12: melting point 220-222 ° C (EtOH); [α] ²² _D +80.
6 ° (c 0.57, CHCl ₃ ); IR band (KBr) 3350, 1770, 168
0, 1550 cm ^-1 ; ¹ H-NMR (400 MHz, py-d ₅ ) δ 1.47 (3 H,
s, H-14), 1.92 (3H, brs, H-15), 4.32 (1H,
t, J 10.5Hz, H-6), 5.42 (1H, brs, H-2),
5.50 (1H, br s, H-3), 5.41 (1H, d, J 3.5Hz,
H-13a), 6.15 (1H, d, J 3.5Hz, H-13b).

Cooled solution of 120 mg of diol 10 in pyridine (about 0 ° C)
Then, about 0.1 ml of POCl ₃ was added to synthesize epiarglabin 13, which is the 1,10-epimer of algrabin (FIG. 2).
After stirring at about −5 ° C. for 24 hours, the reaction solution was extracted with ethyl ethanol to purify the reaction solution (worked up). 5% H
After washing with Cl and water, the residue was crystallized from petroleum ethyl ethanol to give 40 mg of 1,10-epilagrabine 13.

1,10-Epiargrabine 13 Features Overview: Melting Point 193-1
94 ° C (EtOH); [α] ²⁰ _D + 78.4 ° (c 0.43 CHCl ₃ ); IR
Band (KBr) 1760, 1665, 1650, 1150 cm ^-1 ; ¹ H-NMR (4
00MHz, py-d ₅ ) δ1.30 (3H, s, H-14), 1.90 (3
H, brs, H-15), 2.66 (1H, m, H-5), 4.18 (1
H, dd, J 14.5, 12.5Hz, H-6), 5.43 (1H, m, H
-3). 5.38 (1H, m, H-3), 6.14 (1H, d, J 3.
5Hz, H-13b).

The interaction between arglabin 1 and epoxy arglabin 2 by benzenamine, dimethylamine, and morpholine in alcoholic medium is chemoselectively (as a Michael reaction to activated double bonds of these molecules) ( chemoselectively), yielding 56-85% of the corresponding derivatives 14a-d and 15a-d (FIG. 3).
The spectrum revealed that the aminomethyl residue was in the α-coordinate.

Synthesis of dimethylamino arglabin 14b: arglabin 1 was mixed with 0.21 L of alcohol and heated to 40 ° C. until the arglabin was completely dissolved. After filtering with a filter, a 33% solution of dimethylamine (0.023 L) was added dropwise with stirring. The mixture was left at room temperature for 24 hours. The reaction was observed on silufol plates using TLC. After the activation reaction was completed, the mixture was heated to 52 ° C and the alcohol was distilled under reduced pressure. About 0.63 L of chloroform was added to the remaining solvent and stirred for 30 minutes. This mixed solution was poured into a separating funnel, and the portion where chloroform was visible in the lower part of the funnel was recovered. Chloroform extraction was repeated twice more on the aqueous layer. The recovered chloroform was dried using magnesium sulfate. The chloroform-magnesium sulfate mixture was stirred for 30 minutes and then filtered under reduced pressure to remove chloroform. About 22 g of dimethylaminoarglabin 14b was produced.

Dimethylaminoarglabin 14b was first dissolved in 5 volumes (w / v) of chloroform and then about 3 volumes (w / v).
/ v) KCK silica gel. After evaporation of the solvent, the dried material was chromatographed on a KCK silica gel column made up of addition product and adsorbent in a ratio of 1:22. The column was eluted with a mixture of petroleum diether and ethyl ether (1: 1, 1: 2). Fractions of approximately 14-17 ml were collected and observed by TLC. Dimethylaminoarglabin 14b was recrystallized from the chloroform and ether (1: 1) fractions.

An overview of the characteristics of dimethylaminoarglabin 14b: melting point 9
4.5-95.5 ° C; [α] ²¹ _D + 47 ° (c 1.7 CHCl ₃ ); Component analysis 70.41% C, 8.7% H, 4.82% N (C ₁₂ H ₂₅ O ₃ N), I
R band (≧ CHCl max) 3050 to 3000 (shoulder), 2940, 286
0, 2835, 2780, 2410, 1770 (carbonyl lactone) 165
0 (double bond), 1550 to 1530 (wide band), 1470, 145
0, 1385, 1335, 1180, 1150, 1140, 1125 cm ^-1 (epoxy group); MS (m / z,% strength) M + HCl 291 (5.07, HCl),
247 (0.5), 188 (1,2), 115 (2,19), 105 (1,6), 9
7 (3,2), 77 (3,5), 70 (6,2), 67 (2,9), 58 (10
0); NMR (200MHz, CDCl ₃ , δ scale; multiplet, PPM
KCCB) 1.90 (3H), 2.27 (6H), 4.00 (1H, = 9.5), wide singlet 5.53 (1H), dm2.66 (2H, J4 = J2 = 5.5).

Dimethylaminoarglabin 14b was dissolved in 0.22 L of alcohol and heated to 40 ° C. to generate dimethylaminoarglabin hydrochloride 14d. After vacuum filtration, 0.2 kg of sodium chloride and a few drops of concentrated sulfuric acid were added to generate hydrogen chloride gas. The reaction was monitored by TLC. When the reaction was complete, the mixture was heated to 52 ° C and ethanol was distilled under reduced pressure.
To the remaining tar was added about 0.9 L of ethyl acetic acid with vigorous stirring. The resulting precipitate provided about 21 g of dimethylaminoarglabin hydrochloride 14d.

About 0.1 L of chloroform was added to dissolve dimethylaminoarglabin hydrochloride 14d and then distilled to remove chloroform. The remaining tar was mixed with 0.83 L of ethyl acetate with vigorous stirring. The mixture was cooled to ensure the product was completely precipitated.
The resulting precipitate was vacuum filtered to remove all solvent. The final product was dried under vacuum to anhydrone and dissolved in athermal distilled water at a ratio of 100 g water to 2 g dry material. The yield of dimethylaminoarglabin hydrochloride 14d was about 20 g (95% of the estimated amount at this stage).

Summary of the features of dimethylaminoarglabin hydrochloride 14d:
Melting point 203-204 ° C (ethanol ether); [α] ²¹ _D +6
1.53 ° (c 0.52, CHCl ₃ ); IR band 33050-3000 (broad band), 2980, 2970 (broad band of intensity, NH), 28
90, 2970, 2360-2300 (wide band), 1775 (carbonyl group of lactone), 1650 (thin band), 1480, 1450,
1385, 1345, 1185, 1140-1120, 1100, 1065, 1040, 10
10cm ^-1 ; MS (m / z,% strength) 291 (3.01, M ⁺ HCl), 115
(2.19), 105 (1.5), 97 (3.2), 91 (4.0), 77 (3.
5), 70 (16.2), 67 (2.9), 58 (100); NMR (200MH
z, CDCl ₃ , δ scale; multiplet, PPMKCCB) c.1.30
(3H), c.1.87 (3H), c.2.87 (6H), dm4.17 (1H,
J1 = J2 = 10Hz), wide singlet 5.55 (1H).

Treatment of Algravin 1 with ethanol and H ₂ / Ni, 11,
13 Dihydroarglabin 16 was produced.

Example 3: Freeze-drying-an aqueous solution of dimethylaminoarglabin hydrochloride was passed through a cotton gauze stopper or a stack of 8 pieces of gauze and then filtered through a sterile Millipore filter to obtain a sterile glass bottle. I put it inside. This solution was sucked into a measuring buret with a vacuum pump and dispensed into 2 ml vials or ampoules in equal amounts. The vial or ampoule containing the solution is KC-30
Stored in sterilized shelves at −40 ° C. for 24 hours before being dried in freeze dryer or LS-45 freeze dryer. After this preparation period, the drying process was started. After maintaining the temperature at -40 ° C for 2 hours, the temperature was gradually raised to about 50 ° C (plus or minus about 5 ° C). The change to about 50 ° C. was allowed to occur over about 12 to 13 hours. The final temperature did not exceed +60.
The total drying time was 24 hours. After this,
The vial containing the dried compound was immediately capped and rotated. The ampoule was sealed. Each vial or ampoule contained approximately 0.04 g of preparation.

Vials or Ampoules Filters Non-sterilized vials or ampoules were sterilized by autoclaving at 120 ° C for 20 minutes at 1.2 atmospheres.

Alternatively, the prepared dimethylaminoarglabine hydrochloride aqueous solution was filtered through a cotton gauze stopper or an 8-layer gauze. About 200 ml of the solution was poured into a 500 ml bottle, covered with a cotton gauze stopper or an eight-layer gauze, and wrapped with oil paper. The bottle containing the aqueous solution was sterilized by autoclaving at 120 ° C. for 30 minutes at 1.2 atm. The sterilized solution was cooled to room temperature. Using a sterilization technique, 2 ml of the solution was poured into a 10 ml vial.
The vial was then lyophilized as described above. After lyophilization, each vial contained approximately 0.04 g of preparation.

The yield of this compound is 17 g, which corresponds to 8% of the material at this stage.
Equal to 8.2%, 0.22% of all natural dry materials. The lyophilized material had a whitish straw color and had a bitter taste. That this preparation is authentic,
Melting points were measured and confirmed by recording IR-spectra, mass spectra and NMR spectra. The quality of the preparation was checked by diluting 1 mg of preparation with 0.2 ml of water. Upon the addition of a drop of vanillin solution saturated in concentrated sulfuric acid, the mixture turned purple indicating the presence of terpenes.
The lyophilized material can be stored for 3 years.

Example 4: Isolation of Other Sesquiterpene Lactones The structures of compounds 17-21 are shown in FIG. Gravelin 17
Also, Artemisia glavella Kar.et Kir.
labella Kar.et Kir.). The yield from dried raw material was about 0.016%. The structure of Gravelin was determined by IR-spectrum, UN-spectrum, NMR spectrum, C13 NMR spectrum, mass spectrum, and chemical transition.

An overview of the characteristics of Gravelin: melting point 130-131 ° C (petroleum diether); [α] ²⁰ _D + 90.9 ° (SO, 17, chloroform).

3-keto-eudesm-1 was obtained by selective dehydration of α-sautonine in 45% yield.
(2), 4 (5), 11 (13) -triene 6,12-olid (1) 18 was prepared, which can be prepared from 20 or more types of mugwort. This structure is
Determined by UV-spectrum and NMR spectrum.

Summary of 18 features: melting point 145-147 ° C (methanol);
[Α] ¹⁸ _D -10.4 ° (with 1,12; chloroform).

Anovin 19 is Achilles nobilis L. (Achilles nob
ilis L.). 2α, 3α-epoxy-4α, 10α-dioxy-5,7α (H), 6β (H) -of Anovine 19
The guay-11 (13) -ene-6,12-olide structure was confirmed by IR-spectra, NMR spectra, mass spectra and chemical transitions.

Epoxy estafiatone is produced by isomerization of trifluoride boron with etherate to isomerize available terpene lactones such as estafiatine, followed by epoxidation with richlorobenzoyl acid. 20
(Epoxy estafiaton) was produced. 3-keto-10α (1
4) -Epoxy-1,5,7α (H) 4,6β (H) -Guay-1
The 1 (13) -ene-6,12-olide structure was determined by IR-spectra, NMR-spectra and mass-spectra.

Gaillardio grandif
lora) is extracted with chloroform and then separated by chromatography on a silica gel column,
Geigranin 21 was produced. The structure of geigranin 21
It was determined by IR-spectrum, UV-spectrum, NMR spectrum and Cesy-spectrum.

Example 5: In vitro activity of arglabin, and its derivatives-cell viability-Primary cultures of transformed and normal cells are incubated with various concentrations of dimethylaminoarglabin hydrochloride to affect cell viability. The effect was measured.

Mouse mastocytoma (P-815), and myeloma (Z
-P3X63Ag8.653 and Pai) cell lines, as well as the human erythroleukemia (K-562) cell line. Normal mouse hepatocytes were isolated from mouse liver using collagenase. Mouse splenocytes were isolated using a glass homogenizer. Bone marrow was washed to obtain bone marrow cells. For example, Shears (SB) and Kirk (CJ), (198
4), Biochem. J. 219: 375-382.

RPMI-supplemented with 10% fetal bovine serum, 100 mM L-glutamine, and 50 μg / ml gentamicin.
The cells were cultured in 1640 medium at 37 ° C. under 5% CO ₂ . Cells were seeded in 24-well plates at a density of 50,000 cells per well and allowed to grow for approximately 2 days to near confluence before being transferred to 96-well plates at the same density. The transformed cell lines were incubated with dimethylaminoarglabin hydrochloride in the concentration range of 1.5 μg / ml to 100 μg / ml.
Cell viability was measured by trypan blue fractionation. As shown in FIG. 5, it was 6 μg / ml for X-653 cells and K-562 cells, and 1 for P-815 cells.
At 2 μg / ml, viability was reduced by half. At a concentration of 12 μg / ml, approximately 25% of K-562 cells and X-653 cells survive and
At a concentration of 25 μg / ml, the same proportion of P-815 cells survived. With the highest concentration of dimethylaminoarglabine hydrochloride,
The viability of all transformed cells was further reduced.

Proliferation of transformed cells was measured by incubating ³ H thymidine in medium for 18 hours. After a lapse of a predetermined time, the amount of ³ H thymidine incorporated was measured to measure proliferation. Thymidine incorporation provides a quantitative measure of DNA synthesis rate, which is generally directly proportional to cell division rate. FIG. 6 shows X-653 cells and P.
The proliferation of −815 cells was effectively inhibited at the concentrations of 6 μg / ml and 12 μg / ml, respectively.

Primary culture of normal cells from 10 μg / ml to 2560 μg / ml
With dimethylaminoargrabine hydrochloride in a concentration range of 18
Incubated for hours. Cell viability was measured by trypan blue fractionation. FIG. 7 shows that increasing the concentration of dimethylaminoarglabin hydrochloride reduces the viability of normal cells, but that compared to transformed cells, a considerably higher concentration is required to kill normal cells. Shows. At a concentration of 320 μg / ml, the number of splenocytes was 50% less than in controls. At concentrations of 640 μg / ml and 1280 μg / ml, 40% and 10% of splenocytes were still alive, respectively. At these same concentrations, about 50% and 25% of hepatocytes were still alive, respectively. Bone marrow cells were more sensitive to dimethylaminoarglabin hydrochloride. 160 μg / m
At a concentration of l, only about 50% of bone marrow cells survived.
When the concentration was raised to 320 μg / ml, the viability dropped to about 25%.

Protein prenylation—Pai cells, which are mouse bone marrow cells, were cultured in the presence of 60 μM dimethylaminoarglabin hydrochloride. The cells were harvested by centrifugation at 600 xg for 10 minutes and then washed twice with PBS. Control cells were cultured without drug. Lysis buffer (50 mM Tris, pH 7.4, 25 mM EDTA, 0.05% Tween, 0.01 M Na for 30 minutes on ice
The cells were solubilized in Cl). A cell lysate was prepared by homogenizing at 4 ° C. for 5 minutes, and centrifuged at 12,000 × g for 10 minutes to precipitate. The supernatant was collected. After the protein was precipitated with trichloroacetic acid, it was washed successively with ethanol and ethyl ether. Selective naphthol cleavage of the bond between the isoprenoid and the protein is reported by Epstein (Ep
(stein, WW) et al. (1991) Proc.Natl.Acad.Sci.USA 88:
It was carried out as described in 9668-9670. Generally, potassium naphthoxide (potassuim naphthoxid)
5 mg of a 4: 1 mixture of e) and naphthol was added to approximately 10 mg of precipitated protein. After adding 50 μl of dimethylformamide, put argon gas into the tester and cover with a lid.
Heat to 100 ° C. for ˜15 hours. The reaction product was extracted with hexane and using a 0.4 × 15 cm reverse phase Nova-Pac C ₁₈ column,
Analysis by HPLC (Waters System) was performed. The column was eluted with 20% water in acetonitrile at a flow rate of 1.0 ml / min. The naphthol cleavage product was detected at 360 nm with an actual refraction of 0.01 A units (Figure 8). In the control (FIG. 8A), the farnesyl cysteine derivative was eluted at 4.5 minutes and the geraniugeranyl cysteine derivative was eluted at 6 minutes. The molar ratio of geranylgeranyl to farhesyl cysteine was 6.

The effect of dimethylaminoarglabine hydrochloride on cell prenylation is shown in Figure 8B. When 60 μM of dimethylaminoarglabin hydrochloride was used, the chromatogram did not show a peak corresponding to the farnesyl cysteine derivative, but the peak of geranylgeranyl cysteine derivative appeared as in the control. This is
Dimethylaminoarglabin hydrochloride has been shown to interfere with protein farnesylation without significantly affecting geranylgeranylation.

Example 6: In vivo activity of arglabin, and its derivatives Overall, proteins of this family are less toxic and tolerated at supra-therapeutic doses. Dimethyl sulfoxide (DMSO) using traditional toxicological methods
LD ₅₀ was measured by intraperitoneally injecting a 2% dimethylaminoarglabine hydrochloride solution into mice (body weight 20 to 22 g) and rats (body weight 120 to 130 g). LD ₅₀ is 190-220 mg / k in mice
g, and in rats, it was 280-310 mg / kg. At autopsy of the animals, visceral polycythemia, ie mesenteric and intestinal vasodilation, was clearly seen.

In rats and rabbits, the doses did not impair liver, kidney, cardiovascular, respiratory, or peripheral nervous system functions. Blood pressure was unchanged.
Furthermore, no febrile, allergic, teratogenic, or embryotoxic effects were seen in animals.

Rats, guinea pigs, or mice were given ip and rabbits were given iv daily to determine the maximum tolerated dose (MTD) of algrabin and its derivatives for 5 to 20 days. Generally, for all compounds tested, the MTD is from about 20 mg / kg to about
For example, the maximum dose of dimethylaminoarglabine hydrochloride in DMSO solution, which ranged up to 50 mg / kg, was
From 20mg / kg to 30mg / kg in mice, 45mg / k in guinea pigs
g, 50 mg / kg of rat. When the intraperitoneal administration of a 2% aqueous solution of dimethylaminoarglabine hydrochloride was continued for a prolonged period of time, reversible changes in glycolysis and respiratory action of tissues were observed in blood serum and liver tissues, and changes in hormone balance were observed. , And an increase in the amount of protein in urine was observed.

Tumor Growth Inhibition in Rats-Transplantation of human tumors into mice and rats (sarcoma M-1; plis lymphosarcoma; carcinosarcoma of worker); gelen (Ge
ren) carcinoma; sarcoma 45; sarcoma 180; sarcoma 37; liver alveolar carcinoma (Alveo
PC-1; Ehrlich's solid adenocarcinoma of Ehrlich; Breast cancer (PMK);
Lymphocytic leukemia P-388; lymphoid leukemia L-1210; rubidomycin, prospidine, and leukoeffdine-resistant mutants of plys lymphosarcoma; Resistant sarcoma 45 variants). After transplanting to mice
The treatment was started 24 hours later, and in rats, the treatment was started when a measurable tumor node was detected. The control animals are
It consisted of a county of 10 to 15 animals. To assess the compound's anti-tumor activity, the percentage of tumor growth inhibition was determined after termination of treatment. The results were statistically analyzed using t-test. Histologically, tumor regression was associated with dystrophies, necrosis of tumor cells, inhibition of blood supply to tumor tissue, and replacement with connective tissue.

Tables III and IV summarize the percent tumor growth inhibitory activity of algrabin and its various derivatives against both drug-resistant and drug-resistant tumors. For comparison, Table III also includes the percent tumor growth inhibition for colchicine of compounds known to have antitumor activity. Epoxidation of algrabin on the C3-C4 double bond also increased antitumor activity. Dimethylaminoarglabin and dimethylaminoarglabin hydrochloride were effective against a wide range of tumors. One of the advantages of dimethylaminoarglabin hydrochloride is that it is water soluble.

Combined therapy-animal studies have allowed the design of the most rational treatment regimen with dimethylaminoarglabine hydrochloride and other anti-tumor agents.

Of rats treated with a combination of dimethylaminoarglabin hydrochloride, cisplatin, and methotrexate
At 60%, complete loss of tumor resistance to prospidin and rubidomycin was observed. In addition, this combination allows for sarcoma-45 methotrexate,
It was possible to overcome the cross-resistance of sarcoma-45 to 5-fluorouracil and to pris lymphosarcoma to rubidomycin. No animals died from this treatment.

Leukofedin after sarcolicin administration
The complete disappearance of the tumor was observed in 60% of the rats, while the resistant plis lymphosarcoma was secondarily susceptible. The combination of dimethylaminoarglabine hydrochloride and sarcolicin resulted in inhibition of DNA synthesis at half the maximum tolerated dose (MTD) (synthesis inhibition index 94.1-97.1%). No reduction in blood cell mass occurred with this combination.

By combining dimethylaminoarglabine hydrochloride and methylnitrosourea, the administration interval of these two drugs is 2,
It was administered at 4 or 24 hours. It was determined to be optimal to administer dimethylaminoarglabine hydrochloride 2 hours prior to administration of methylnitrosourea. Sarcoma −45
Of sarcoma-45 against 5-fluorouracil of sarcoma-45, rubidomycin of plis lymphosarcoma, and of prospidin of pris lymphosarcoma were combined with dimethylaminoarglabin hydrochloride and methylnitrosourea. About 60% of the tumors disappeared in rats without adverse drug reactions.
Administration of methylnitrosourea before administration of dimethylaminoarglabin hydrochloride reduced antitumor activity and increased toxicity.

Histologically, the number of small nuclear enriched polymorphic cells with no clear structure was smaller than that of the control group after the combined treatment with dimethylaminoarglabine hydrochloride and methylnitrosourea. It was found that administration of dimethylaminoarglabine hydrochloride 2 hours before administration of methylnitrosourea reduced toxicity.

The same result was seen when administering dimethylaminoarglabine hydrochloride 2 hours before the administration of the vincristine and vinblastine mixture for gellen carcinoma and worker sarcoma.

Dimethylaminoarglabin hydrochloride slightly prolonged the survival of animals with non-resistant and drug resistant tumors. The combination of dimethylaminoarglabine hydrochloride with other antitumor agents further extended survival. For example, the combination of dimethylaminoarglabine hydrochloride and vincristine increased the survival of animals bearing methylnitrosourea-resistant tumors by 114%. The combination of dimethylaminoarglabine hydrochloride and cisplatin increased the survival of animals with methotrexate-resistant L1210 by 117% at half the MTD. When three types of dimethylaminoarglabin hydrochloride, vincristine, and cyclophosphamide are combined and half the MTD is administered, when dimethylaminoarglabine hydrochloride and vincristine, or dimethylaminoalgrabin hydrochloride and cyclophosphamide are combined Better effect was seen. Survival time of 209% due to the combination of 3 types
It became longer. The combination of dimethylaminoarglabin hydrochloride, vincristine, cyclophosphamide, and cisplatin reduced the effect more than the combination of three. This may be due to the increased toxicity of antitumor agents.

The effects of dimethylaminoarglabin hydrochloride alone and its effects in combination with other drugs were investigated in a drug-resistant metastasis model of plis lymphosarcoma. Of the primary drug-resistant lymph nodes, metastases in inguinal lymph nodes were the most sensitive. These did not occur in 10% of cases. With dimethylaminoarglabin hydrochloride alone, the survival time was 128% of the control group. The combination of dimethylaminoarglabine hydrochloride and vincristine resulted in tumor destruction and inhibition of tumor growth in 30% of rats. No new metastases in inguinal lymph nodes, survival 174%
Extended.

The combination of dimethylaminoarglabine hydrochloride and methotrexate resulted in 300% longer survival in animals with prospidin-resistant plis lymphosarcoma. This combination resulted in an 8-fold lower metastasis frequency.

In order to clarify the possible mechanism of action of dimethylaminoarglabine hydrochloride on primary drug-resistant tumors and their metastasis, sarcoma 45 And used to treat inhibition of DNA synthesis. In the case of drug-resistant sarcoma 45, beneficial results were observed with sarcosyl and the combination of sarcosyl and dimethylaminoarglabine hydrochloride. Drug resistant sarcoma
In the case of 45, dimethylaminoarglabine hydrochloride alone was very effective (DNA inhibition index 99%). In addition, DNA inhibition was increased 24 hours after daily administration for 5 and 10 days. This indicated that repeated administration of dimethylaminoarglabine hydrochloride accumulated an anti-tumor effect as compared to administration of MTD at once.

After treatment with dimethylaminoarglabine hydrochloride alone and in combination with another cytostatic agent, immunity of rats with metastatic primary prospidin-resistant tumors was investigated. Treatment of animal tumors with dimethylaminoarglabin hydrochloride was observed to ameliorate the immunosuppression seen after treatment with sarcolicin and cisplatin. Administration of dimethylaminoarglabine hydrochloride in combination with sarcolicin or cisplatin, especially dimethylaminoarglabine hydrochloride 2 hours before administration of cytostatics, increased the immunological index. These results suggested that dimethylaminoarglabine hydrochloride moderated the immunosuppressive effects of cytostatics and normalized the body's immune balance. These data indicate that dimethylaminoarglabine hydrochloride, alone or in combination, reduced cytotoxicity and enhanced efficacy against drug resistant tumors.

Modulation of the immune system-Effects of dimethylaminoarglabin hydrochloride were measured in naive or immunocompromised mice. Mice were immunosuppressed by administration of 200 mg / kg of cyclophosphamide. When you inject cyclophosphamide,
Significant leukopenia occurred, mainly associated with lymphopenia. The animal's humoral immunity was significantly suppressed and, to a lesser extent, cell-mediated immunity. A 2% dimethylaminoarglabin hydrochloride solution was intraperitoneally injected into white hybrid mice at two doses of 50 mg / kg and 100 mg / kg. The hemagglutination test and delayed hypersensitivity reaction were measured before and after drug administration. 1 dimethylaminoargrabine hydrochloride
It was determined that hemagglutination concentration or delayed type hypersensitivity reaction was not changed by single dose of 50 mg / kg. 100 mg / kg
Was slightly decreased in hemagglutination concentration.

10 to 50 mg / kg of dimethylaminoarglabine hydrochloride was repeatedly intraperitoneally injected, administered for 5 to 10 days, and the effect of prolonging the administration period was measured. With doses as low as 10 mg / kg and 20 mg / kg,
Hemagglutination concentration increased from the 5th day to the 10th day. 30 mg
By administration of higher doses such as / kg, by day 10,
The result was that the hemagglutination test index decreased. No effect on delayed type hypersensitivity was seen on day 5, but 10
It had risen by day one.

In untreated mice, injection of 10 mg / kg increased the absolute number of lymphocytes and thus the total number of white blood cells.
The relative number of neutrophils decreased slightly. 20 mg / kg
However, there was no overall increase in white blood cell count. Using the nitroblue tetrazolium assay,
The function of neutrophils was evaluated. At 10 mg / kg, the number of neutrophils decreased, but there was no change in neutrophil activity. Dose
At 20 mg / kg, there was a decrease in nitroblue tetrazolium-positive neutrophils, which was indicative of reduced function.

Intraperitoneal administration of 10 mg / kg and 20 mg / kg of dimethylaminoarglabin hydrochloride daily for 10 days resulted in dramatic changes in T lymphocytes, B lymphocytes, and natural killer cells. At 10 mg / kg, the increase in natural killer cells and T lymphocytes increased the overall white blood cell count, while B lymphocytes remained stable. The increase in the number of T lymphocytes was found to be the result of an increase in the amount of T helper subgroups. The amount of T suppressor subgroup was unchanged. Large dose (20mg
/ kg), the number of B lymphocytes and T lymphocytes also decreased to a lesser extent, but the total white blood cell count did not change. The number of natural killer cells was increasing. It was also reduced in the nitroblue tetrazolium assay.

Overall, the effect of dimethylaminoarglabin hydrochloride on the immune system was dose-dependent. Low dose (10 mg / k
In g), with dimethylaminoarglabine hydrochloride,
There was an increase in the amount of T lymphocytes, B lymphocytes, and natural killer cells. The increase of T lymphocytes is
It was accompanied by an increase in the amount of helper subgroups. At the higher dose of dimethylaminoarglabine hydrochloride (20 mg / kg), T
The number of lymphocytes and B lymphocytes decreased, but another constant cell population, such as natural killer cells, increased.

Dimethylaminoarglabine hydrochloride in immunocompromised mice
Injection of 10 mg / kg for 10 days was observed to reduce leukopenia and lymphopenia in mice. By day 10 of treatment, the total number of lymphocytes in immunosuppressed animals was unchanged from that obtained from untreated animals. The increase in T lymphocyte counts was slightly accompanied by an increase in T helper and T suppressor subgroups. The number of B lymphocytes is completely
It was not restored to normal.

Further immunological data were obtained from rats of the "August" strain of rats, weighing 140-160 g, which may or may not have plis lymphosarcoma. Spontaneous erythrocyte rosette formation, nitroblue tetrazolium assay, delayed hypersensitivity reaction, and hemagglutination, before, during (5 and 10 days), and 5 days after treatment with dimethylaminoarglabin hydrochloride. I examined one index. 2 of lyophilized dimethylaminoarglabine hydrochloride
% Aqueous solution of about 50 mg / kg was intraperitoneally injected daily for 10 days.
The results are summarized in Table V. In summary, lyophilized dimethylaminoarglabine hydrochloride stimulated delayed type hypersensitivity reactions in untreated rats, but reduced other investigated indices. In rats with prislymphosarcoma all immune system parameters were elevated and the immune system was stimulated. The advantage of lyophilized dimethylaminoarglabine hydrochloride is 5
-To improve the immunosuppressive effect of cytostatics such as fluorouracil and sarcolicin.

Pharmacokinetics-Using 30 male and female randomized rats,
The pharmacokinetic data of dimethylaminoarglabin hydrochloride were collected experimentally. Rats weighed 200-220 grams. Gas chromatographic and FARM modeling programs were used in the analysis of all pharmacokinetic data. When 2 mg / kg of dimethylaminoarglabine hydrochloride is put into the vein, the maximum amount in serum is 30 μg / ml in 1 hour.
showed that. Dimethylaminoarglabin hydrochloride rapidly spread throughout the body of organisms, from blood to peripheral tissues. Due to the large apparent distribution capacity, this is
It has been shown to be able to cross cell membranes and tissue barriers. For the first hour after administration, the maximum drug concentration is
It accumulated in the lungs and spleen. The highest concentrations in lung and spleen were 149.4 and 159 μg / g, respectively. Within 3 hours after administration, the concentration in the liver and skeletal muscle was 22
The values were 8.6 and 176.4 μg / g. The preparation accumulates in the liver,
It was found to be retained for a longer period than other organs. Dimethylaminoarglabin hydrochloride was able to penetrate the blood-brain barrier. The concentration in brain tissue became 23.9 μg / g after 1 hour, and stabilized at 15.6 μg / g in 24 hours.

Dimethylaminoargrabine hydrochloride was discharged very slowly. The biological half-life was about 46.8 hours in rats with an average residence time of about 67 hours. Excretion from the kidney progressed slowly. The renal concentration reached a maximum after 3 hours. Maximum renal concentration of 56.6 μg / g by 24 hours
Became. The transport rate of the preparation from peripheral tissues to blood was low with a total clearance of 0.05 ml / min.

Clinical Data on Dimethylaminoarglabine Hydrochloride A first clinical trial of dimethylaminoarglabine hydrochloride was conducted on 51 patients with end-stage cancer (III-IV). In the first clinical trial, 20.7% of patients had lung cancer, 17%
Had liver cancer, 17% had gastric cancer, 9.4% had rectal cancer, 5.7% had ovarian cancer, 5.7% had esophageal cancer, and the rest had salivary gland cancer, lymphosarcoma, breast cancer, or colon cancer. 67.9 of the patient population
% Were men and 32.1% were women. In general, dimethylaminoarglabine hydrochloride reconstituted in aqueous solution was administered intravenously to patients. For patients with ascites, dimethylaminoarglabin hydrochloride was administered intraperitoneally. Patients with pleurisy received intrapleural administration. Before proceeding with the study, very small doses were given to the patients and observed for signs of an allergic reaction. First, 80 mg of the compound was administered daily as a single dose, and then gradually increased to the maximum dose. After 30-35 days, the dose is 1
It was increased to 480 mg per day. At such high doses, the patient complained of nausea and vomiting. In a typical case,
It was determined that the daily dose should be approximately 240-280 mg.
The total dose administered during the entire course of treatment was typically 5-6 g of dimethylaminoarglabine hydrochloride, although doses as high as 20 g were also possible. Immediately after the administration of the compound, the patient complained of an immediate disappearing bitterness. Some patients had additional courses of treatment. The data obtained from the first clinical trial are summarized in Table VI. The pre-treatment patient's symptoms were assessed on a scale of 1 to 3, with 1 being bedridden, 2 being fairly restricted in activity, and 3 representing maintaining full activity. The treatment result was measured from 0 to 3. here,
0 is no improvement, 1 is not significant or 1
Improved for a period of less than a month, 2 is significantly improved (25-50% reduction in tumor size) and 3 shows a sharp improvement (50-100% reduction in tumor size).

Collectively, about 30% of patients had a significant improvement. 55.6% of liver cancer patients improved significantly after treatment with dimethylaminoarglabine hydrochloride. Lung and ovarian cancer patients responded particularly well to dimethylaminoarglabine hydrochloride, with 64% and 66% of ovarian cancer patients improving considerably. Dimethylaminoarglabin hydrochloride had little toxicity and did not suppress hematopoietic effects. No negative reactions in the gastrointestinal tract or pores were recorded during the study.

In patients with primary hepatocellular carcinoma, treatment with lyophilized dimethylaminoarglabine hydrochloride resulted in liver shrinkage of more than 50% in two patients, and about 2% in the other.
Reduced by 50%. Patients reported improved mental status and appetite. The pain in the lower right rib disappeared.

The immune status of the patients was evaluated using standard methods of rosette formation and phagocytosis. These indicators were examined before, during and after treatment. Blood samples were taken from the fingers.
Analysis of the mean immunity values for this patient group revealed a positive response to treatment. On days 3-5 of treatment, the number of T lymphocytes was reduced from 57% to 40.1%, the number of T helper lymphocytes was reduced from 50% to 37.3%, and the neutrophil adhesion was 42% to 28.5%. % Has been reduced. Undifferentiated lymphocytes increased from 21.5% to 42.2%. The general change in the ratio of T helper lymphocytes to T suppressor lymphocytes is due to the increase in T suppressor lymphocytes. B lymphocyte counts and phagocytic activity remained stable. The total number of lymphocytes increased to 9.4 x 10 ⁹ / L,
The total number of lymphocytes increased as well. Elevated levels of all immunoglobulin types.

By day 20 of treatment, all indicators had returned to normal. In some patients, the index returned to normal by day 14. In the patients analyzed 30 days after the treatment, an increase in the number of T lymphocytes and an increase in the adhesiveness of neutrophils were significantly observed.

The first clinical trial was discontinued because the production of dimethylaminoarglabin hydrochloride was delayed by 3-6 months. In a second clinical trial, 72 patients (61.1% male, 38.9% female) with variously localized stage IV cancers received dimethylaminoarglabine hydrochloride. Of the patients, 25% had gastric cancer, 16.7% had liver cancer, 18.1% had lung cancer, and the remaining 40.2% had esophageal, breast, ovarian, pancreatic, brain, or lymphosarcoma. I was sick.
Patients in poor condition had liver (25%), retroperitoneal lymph nodes (25%), ascites (22.2%), and exudative pleurisy (1
1.1%) had metastases. Some of these patients had been treated with dimethylaminoarglabine hydrochloride in their first clinical trial. The results of the second clinical trial are summarized in Table VII.

There are many advantages to using dimethylaminoarglabine hydrochloride as an anti-tumor cytostatic agent in solid tumors. This preparation has no side effects, does not suppress the hematopoietic action, normalizes the functional state of the immune system, and has no allergic effect. The ulcer cell growth inhibitor is particularly effective against primary cancers of the liver and other solid cancers associated with polyserositis. Partial tumor regression was observed in 61.1% of cases and the course was stable in 31.9%. Only 7.0% had relapses. 88.9% of patients
(64 of 72) responded to treatment, but 11.1% did not (8 of 72).

The following is a summary of case records of patients selected from those who received dimethylaminoarglabine hydrochloride monotherapy.

Patient M, 55 years old, case number 305, was hospitalized because of numerous nodules on the ribcage and abdominal skin, an ulcer in the excised breast, and hardening of the right chest. Previously, for breast cancer, Sakhalinsk Cancer Center
A mastectomy was performed and a radical mastectomy was performed. After surgery, she received 6 courses of multidrug chemotherapy with cyclophosphamide and methotrexate.

As the illness recurred, symptomatic treatment was recommended. Prior to treatment, there were numerous metastatic nodules ranging from 0.5 to 1 cm on the skin of the abdomen and thorax. On the left side of the rib cage, an ulcer surface of about 10 x 12 cm appeared. The right breast was deformed by infiltrating metastases. Edema appeared in the lower limbs.

The following parameters were known by pretreatment blood analysis. Hb-89, ESR-6mm / hy, L-3.3, Er-3.8m
l, juv.ne-4, seg.ne-78, mon-1. This patient received 5 courses of dimethylaminoarglabine hydrochloride treatment and the total dose was 6.0-7.3 g. A second blood analysis after chemotherapy revealed the following parameters: Hb-122, ESR-20mm / h, L-10.9, Er-3.8ml, eos
-1, stab ne-3, seg, ne-64, lym-34, mon-2.

During the course of treatment, the ulcer became epithelial, the metastatic nodules disappeared and the right breast infiltration was reduced by 50%. Edema of the lower extremities also disappeared.

A 4-month-old patient, case number N2225, and complete cure of liver cancer were seen. This young patient was diagnosed with fetal cancer of the liver with very serious symptoms and was admitted to surgery at the Karaganda Cancer Treatment Center. The rind of the skin was yellow. The patient was breathing painfully. Heart sounds were clear and rhythmic. It was Ps-150 per minute. The tongue was damp and clean. The abdomen was swollen. The liver is swollen and is the lower rim of the bosh-shaped area above the ilium. Hardened with a smooth surface.

Ultrasound tomography (UST) of the liver showed that the liver was swollen and occupied the entire abdominal cavity. The structure was dissimilar because it was a heterogeneous lesion with a hydrophilic edge of up to 5-6 cm, indicating liver cancer.

Pretreatment blood analysis revealed the following parameters: Hb-84, ESR-4mm / h, L-10.9, Er-3.3m
l, eos-1, juv, ne−55, stab ne−45, seg, ne−14, l
ym-30, mon-5. The liver was opened under UST control.
Bare nuclei of tumor cells were observed against a background of degenerated hepatocytes. This patient was diagnosed with fetal liver cancer.

The dimethylaminoarglabine hydrochloride treatment course began with a daily dose of 120 mg IV. The total dose for this course of treatment was 2040 mg. Significant improvement was seen during the course of treatment. Due to the reduced size of the liver, the abdomen became symmetric and smaller.

By UST, at 4 cm along the central rib line, the liver is protruding from under the rib arch, the contour is uniform, and the structure is heterogeneous due to the center of the heterogeneous structure with hydrophilic edges, In addition, the diameter of lesions with high echogenicity is 2.0 to 2.5.
It was shown to be cm. Conclusion: A tumor with focal changes.

A blood analysis again after treatment revealed the following parameters: Hb-177, ESR-4mm / h, er.-
4.0 ml, Z-9.8, eos-3, seg.ne-28, lym-53, mon
-6. The child has been released to good condition. Two weeks later, the course of treatment was repeated, which was well tolerated by the patient.

As of the beginning of 1997, this baby is in good condition. The mother of this child has reported no signs of recurrence. On palpation of the abdomen, the liver is smooth and below the margin of the rib arch.
I found it protruding from 2 cm. This baby is considered cured.

Patient A, 27 years old, case number 543, was diagnosed with a brain tumor. The neurosurgeon determined that there was no possibility of surgery because of the poor condition of the patient. The patient was very weak and had akinetic slowness of the akineticorigid syndrome type. Left and right eye protrusions were reported. After the first course of treatment with dimethylaminoarglabine hydrochloride, the symptoms became stable and he did not complain of headache. After the second course the symptoms were stable. He stopped complaining of headaches and remained appetite. By this case,
Confirmed experimental findings regarding the ability of this compound to cross the blood-brain barrier.

Karnofsky Indicator, 1997 (Table VII
According to I), the efficacy of treatment with dimethylaminoarglabin hydrochloride alone was evaluated. No side effects of dimethylaminoarglabine hydrochloride treatment were reported. The average index of peripheral blood is shown in Table IX.

Immune system indicators were measured using the rosette formation method and phagocytosis. Took dimethylaminoarglabin hydrochloride
57 patients were examined. In order to evaluate the immune status, 11 indices of cell-mediated immunity and humoral immunity were measured for each patient. The following indicators were measured in 0.05 ml of peripheral blood. Absolute and relative amounts of T and B lymphocytes, undifferentiated "zero"
These are adhesion activity and phagocytic activity of cells and neutrophils, hemogram, and serum immunoglobulin amount. Indicators were measured before treatment, on days 2, 5, and 14 of treatment, and after treatment. Table X
Is a summary of the results before and after treatment.

On day 2 or 5 of treatment, the proportion of T lymphocytes and T helper lymphocytes was significantly reduced. The amount of undifferentiated "zero" cells increased. This undifferentiated cell population consisted of mature and undifferentiated B lymphocytes, T lymphocytes, and natural killer cells. No significant change was seen in the hemogram.

From day 6 to day 10 of treatment, almost all indexes returned to the initial values. It was recorded that the proportion of T lymphocytes and their T helper population increased in 2 weeks.
The number of B lymphocytes was reduced. No changes in serum immunoglobulin levels were recorded at this time.

After treatment, a statistically insignificant increase in the proportion of T lymphocyte content was seen. The absolute number of T lymphocytes and the number of T helper lymphocytes that promote the phagocytic effect of neutrophils increased. The number of B lymphocytes increased, and the amounts of immunoglobulins A, M, and G also increased. The number of undifferentiated cells was reduced.

The total number of lymphocytes in the peripheral blood increased. Tumors can cause changes in blood cells both quantitatively and qualitatively, so these parameters were checked after treatment with dimethylaminoarglabine hydrochloride. There was no change in qualitative (morphological) composition of blood cells, but some quantitative changes such as decrease in neutrophil count and increase in lymphocyte count were observed. This suggests a lymphotoxicity reducing effect caused by the tumor. Immunological indices correlated with clinical findings in most cases.

The average value of the immune index was evaluated by the regression analysis method. Integrated indicators such as mean intensity (correlation) expressed in relative units were used to assess the functional status of the immune system. The increase in the index strength of the immune system during the treatment indicates that the immune system responded to the treatment.

Correlation analysis of these data shows that the total amount of true binding parameters increased (r> 0.7 increased binding and negative binding decreased) during treatment.

There was an increased number of interrelationships between immune factors, namely between lymphocyte and neutrophil factors.

Thus, the statistical data analyzed by the different methods of total measurement show that dimethylaminoarglabine hydrochloride is active as a drug that stimulates several immune factors and improves the functional state of the immune system. It indicates that there is. This shows the immunostimulatory effect of this preparation.

Other Embodiments While the present invention has been described in connection with its detailed description, the foregoing description has been made for the purposes of illustration, and limits the scope of the invention as defined by the appended claims. It should be understood that it does not.
Other aspects, advantages, and modifications are within the scope of the following claims.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI C07D 493/14 C07D 493/14 (31) Priority claim number 08 / 934,228 (32) Priority date September 19, 1997 (September 19, 1997) (33) Priority claiming country United States (US) (31) Priority claim number 08 / 934,229 (32) Priority date September 19, 1997 (September 19, 1997) ( 33) Priority claiming United States (US) (31) Priority claiming number 08 / 934,471 (32) Priority date September 19, 1997 (September 19, 1997) (33) Priority claiming United States ( US) (56) References Shaikenov. T. E. , Sensitive effect of Arglabin on transform vs .. normal cells in vitro, Vestnik Ministerstva Nauki--Akademimi Nauk Resplikiki Kazakhstan, Russia, 1996, 6, p55-59 The Merck Index, Co., M. , 1983, tenth edition, No. 2289, 2741, 2815, 4088, 9784, 9788 Ando M. et al. , Studies on the synthesis of sesquiterpene lac tones, 16. The syntheses of 11 <SYM98>, 13-dihydrokaun, J. Mol. Nat. Prod. , UK, 1994, 57/4, p 433-445 Adekenov, S .; M. , Reactions at the double bond and epoxy group of arg labin ", Russian, Khim. Priir. Soedin., Russian, Khim. PRI. Areas (Int.Cl. ⁷ , DB name) C07D 307/93 A61K 31/343 A61K 31/365 A61P 35/00 C07D 493/04 101 C07D 493/14 CA (STN) REGISTRY (STN)

Claims (24)

(57) [Claims] 1. A pharmaceutical composition in unit dosage form suitable for treating human cancer, consisting essentially of about 40 mg to about 480 mg of dimethylaminoarglabine or a pharmaceutically acceptable salt thereof. Pharmaceutical composition. 2. Human cancer includes breast cancer, colon cancer, rectal cancer, gastric cancer, pancreatic cancer, lung cancer, liver cancer, ovarian cancer, leukemia, lymphoma,
The composition according to claim 1, which is selected from the group consisting of pancreatic cancer and esophageal cancer. 3. The composition according to claim 2, wherein the human cancer is selected from the group consisting of lung cancer, liver cancer, and ovarian cancer. 4. The method according to claim 1, wherein dimethylaminoarglabine or a pharmaceutically acceptable salt thereof is freeze-dried.
The composition according to any one of 3 above. 5. The composition of claim 1, wherein the unit dosage form is about 175 mg to about 315 mg dimethylaminoarglabine or a pharmaceutically acceptable salt thereof. 6. The composition of claim 5, wherein the unit dosage form is about 240 mg to about 280 mg of dimethylaminoarglabine or a pharmaceutically acceptable salt thereof. 7. A pharmaceutical composition for treating cancer, which comprises the pharmaceutical composition according to any one of claims 1 to 6. 8. A medicine for treating cancer, which comprises dimethylaminoarglabin or a pharmaceutically acceptable salt thereof. 9. A composition comprising a first chemotherapeutic agent containing dimethylaminoarglabine or a pharmaceutically acceptable salt thereof and a second chemotherapeutic agent for suppressing tumor growth in humans. Composition. 10. The composition according to claim 9, wherein the second chemotherapeutic agent is selected from the group consisting of alkylating agents, antimetabolites, vinca alkaloids, antibiotics and platinum coordination complexes. 11. The alkylating agent is cyclophosphamide,
11. The composition according to claim 10, which is sarcolysin or methylnitrosourea. 12. The composition of claim 10, wherein the antimetabolite is methotrexate or fluorouracil. 13. The composition according to claim 10, wherein the vinca alkaloid is vinblastine or vincristine. 14. The composition of claim 13, wherein the chemotherapeutic agent further comprises cyclophosphamide. 15. The composition according to claim 10, wherein the antibiotic is rubidomycin. 16. The platinum coordination complex is cisplatin.
The composition according to claim 10. 17. A medicament for treating cancer, which comprises the composition according to claim 9. 18. An antitumor agent containing a compound selected from the group represented by the following chemical formulas I, II and III, or a pharmaceutically acceptable salt thereof: In the formula, NRR ₁ is NHCH ₂ Ph or N (CH ₂ CH ₂ ) ₂ O, and N
RR ₂ is NHCH ₂ Ph, N (CH ₂ CH ₂ ) ₂ O, N (CH ₃ ) ₂ or a pharmaceutically acceptable salt thereof. 19. The antitumor agent according to claim 18, which comprises dimethylaminoepoxyarglabin or a pharmaceutically acceptable salt thereof. 20. The antitumor agent according to claim 18, wherein the compound is dibromoarglabin. 21. The antitumor agent according to claim 18, wherein the compound is benzylaminoarglabin. 22. The antitumor agent according to claim 18, wherein the compound is morpholine-aminoarglabin. 23. The antitumor agent according to claim 18, wherein the compound is benzylaminoepoxyarglabin. 24. The antitumor agent according to claim 18, wherein the compound is morpholine-aminoepoxyarglabin.