EP0996441A1 - Pharmaceutical compositions of arglabin and arglabin derivatives - Google Patents
Pharmaceutical compositions of arglabin and arglabin derivativesInfo
- Publication number
- EP0996441A1 EP0996441A1 EP98918509A EP98918509A EP0996441A1 EP 0996441 A1 EP0996441 A1 EP 0996441A1 EP 98918509 A EP98918509 A EP 98918509A EP 98918509 A EP98918509 A EP 98918509A EP 0996441 A1 EP0996441 A1 EP 0996441A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- compound
- arglabin
- composition
- cancer
- treatment
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
- C07D493/04—Ortho-condensed systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/243—Platinum; Compounds thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/34—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide
- A61K31/343—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide condensed with a carbocyclic ring, e.g. coumaran, bufuralol, befunolol, clobenfurol, amiodarone
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/475—Quinolines; Isoquinolines having an indole ring, e.g. yohimbine, reserpine, strychnine, vinblastine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
- A61K31/5375—1,4-Oxazines, e.g. morpholine
- A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/66—Phosphorus compounds
- A61K31/675—Phosphorus compounds having nitrogen as a ring hetero atom, e.g. pyridoxal phosphate
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
- C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D307/93—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems condensed with a ring other than six-membered
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
- C07D493/10—Spiro-condensed systems
Definitions
- RNA synthesis may be inhibited by preventing purine biosynthesis, pyrimidine biosynthesis, the conversion of ribonucieotides to deoxyribonucleotides, antimetabolites, intercalation, or cross-links.
- RNA synthesis may be inhibited by antimetabolites.
- Protein synthesis may be inhibited, for example, by agents that dea inate asparagine. Additionally, agents that inhibit the function of microtubules can be used as chemotherapy agents .
- Chemotherapy agents typically affect both neoplastic and rapidly proliferating cells of normal tissue such as bone marrow, hair follicles and intestinal epithelium. Anorexia, nausea, vomiting, diarrhea, suppression of bone marrow function and hair loss are some of the negative effects commonly associated with chemotherapy. Development of a chemotherapy agent that is an effective antitumor agent with minimal toxicity would be advantageous.
- the invention features a pharmaceutical composition in unit dosage form suitable for the treatment of a human cancer.
- the composition consists essentially of about 40 mg to about 480 mg of arglabin or a derivative thereof.
- the unit dosage of the composition may be, for example, from about 175 mg to about 315 mg or from about 240 mg to about 280 mg of arglabin or a derivative thereof.
- Arglabin or a derivative thereof may be used in the manufacture of a medicament for the treatment of cancer.
- the compositions can be used in the manufacture of a medicament for the treatment of cancer.
- compositions are useful for the treatment of a wide variety of cancers, including, for example, breast, colon, rectal, stomach, pancreatic, lung, liver, ovarian, pancreatic and esophageal cancer, leukemia, and lymphoma.
- the composition is particularly useful for the treatment of lung, liver, and ovarian cancer.
- Dimethylaminoarglabin or a pharmaceutically acceptable salt thereof is a particularly useful arglabin derivative that may be used in the pharmaceutical composition.
- Dimethylaminoarglabin or a pharmaceutically acceptable salt thereof may be lyophilized.
- the invention also features a composition including a first chemotherapeutic agent that includes arglabin or a derivative thereof and a second chemotherapeutic agent.
- the second chemotherapeutic agent is not arglabin or a derivative thereof.
- the composition is effective to suppress tumor growth in a human.
- a particularly useful arglabin derivative is dimethylaminoarglabin or a pharmaceutically acceptable salt thereof.
- the second chemotherapeutic agent may be, for example, an alkylating agent such as cyclophosphamide, sarcolysin, or methylnitrosourea, an antimetabolite such as methotrexate or fluorouracil, a vinca alkaloid such as vinblastine or vincristine, an antibiotic such as rubidomycin or a platinum coordination complex such as cisplatin.
- Two or more chemotherapeutic agents may be in the composition with arglabin or a derivative thereof.
- the compositions can be used in the manufacture of a medicament for the treatment of cancer.
- the invention also features compounds that suppress tumor growth in a mammal . These compounds are selected from the group represented by the following Formulas I, II, III, IV, V and VI:
- RR X is NHCH 2 Ph or N(CH 2 CH 2 ) 2
- RR 2 is NHCH 2 Ph, N(CH 2 CH 2 ) 2 0, N(CH 3 ) 2 , or a pharmaceutically acceptable salt thereof
- X is OH or Cl .
- arglabin derivatives include dimethylaminoepoxyarglabin, dibromoarglabin, arglabin chlorohydrin, 11,13 dihydroarglabin, benzylaminoarglabin, morpholine-aminoarglabin, benzylaminoepoxyarglabin, morpholine-aminoepoxyarglabin, epoxyarglabinchlorohydrin or pharmaceutically acceptable salts thereof.
- Compounds of Formula I, II, III, IV, V or VI can be used in the manufacture of a medicament that suppresses tumor growth in a mammal .
- Figure 2 depicts the synthesis of arglabin derivatives 10 through 13.
- Figure 3 depicts the synthesis of arglabin derivatives 14a-14d, 15a-15d and 16.
- Figure 4 depicts the structure of compounds 17 through 21.
- Figure 5 depicts the effect of increasing concentrations of dimethylaminoarglabin hydrochloride on the viability of transformed cells.
- Figure 6 depicts the effect of increasing concentrations of dimethylaminoarglabin hydrochloride on the proliferation of transformed cells.
- Figure 7 depicts the effect of increasing concentrations of dimethylaminoarglabin hydrochloride on the viability of normal cells.
- Figure 8 is a spectrum index plot of naphthol cleavage products.
- Figure 8A is in the absence of drug.
- Figure 8B is in the presence of drug.
- the invention provides novel compounds that suppress tumor growth in humans .
- These compounds may be synthesized from the parent compound arglabin ( Figure 1) , which is isolated from Artemisia glabella .
- arglabin derivatives may be made using a range of chemistries.
- epoxyarglabin may be produced by epoxidation of the tri-substituted olefin double bond with peracetic acid.
- Dichlorohydrins may be produced by treatment of epoxyarglabin with an ether-acetone HCl solution.
- Dibromoarglabin may be produced by reacting arglabin with Br 2 and carbontetrachloride .
- Arglabin chlorohydrins may be produced from arglabin by reaction with a methanol hydrochloride solution. Epoxidation of arglabin chlorohydrins with peracetic acid and chloroform results in chromatographically separable epoxyarglabin chlorohydrins.
- Arglabin diol, its isomer and diene may be produced by hydrolyzing arglabin.
- the 1, 10 epimer of arglabin, epiarglabin may be produced by treatment of arglabin diol with P0C1 3 .
- Benzylaminoarglabin and benzylaminoepoxyarglabin may be produced by treatment of arglabin and epoxyarglabin with benzeneamine .
- Dimethylaminoarglabin and dimethylaminoepoxyarglabin may be produced by treatment of arglabin and epoxyarglabin with dimethylamine. Morpholine-aminoarglabin and morpholine-aminoepoxyarglabin may be produced by amination of arglabin with morpholine. Pharmaceutically acceptable salts of these compounds may be produced with standard methods and used as antitumor agents. For example, dimethylaminoarglabin hydrochloride and dimethylaminoepoxyarglabin hydrochloride may be produced by hydrochlorination. Dihydroarglabin may be produced by treating arglabin with ethanol and H 2 /Ni. The various arglabin derivatives set out above are depicted in Figures 1-4.
- the invention also relates to a method of suppressing tumor growth in a human patient diagnosed with cancer comprising administering arglabin or a derivative thereof to the patient. While this method may be used generally for the treatment of cancers such as breast, colon, rectal, stomach, pancreatic, lung, liver, ovarian, pancreatic and esophageal cancer, leukemia and lymphomas, certain types of cancers, such as lung, liver and ovarian cancer, are particularly amenable to this therapeutic regimen.
- the compounds can be administered topically, orally, intravenously, intraperitoneally, intrapleurally, intrathecally, subcutaneously, intramuscularly, intranasally, through inhalation or by suppository, depending on the type of cancer and on various patient indications.
- intraperitoneal administration may be used for some patients with ascites.
- Intrapleural administration may be used for certain patients with lung cancer.
- Suppositories may be used for patients with rectal cancer.
- Arglabin or a derivative thereof may be administered in a daily amount from about 40 mg to about 480 mg, preferably from about 175 mg to about 315 mg, more preferably from about 240 mg to about 280 mg.
- the dosage ranges from about 0.5 mg/kg to about 7 mg/kg. In extreme conditions, up to about 20 mg/kg of arglabin or a derivative thereof may be administered. Once administered, these compounds act as antitumor agents and may inhibit the growth of the tumor or may cause the tumor to regress .
- ras gene is a protooncogene that plays a role in many types of human cancers, including colorectal carcinoma, exocrine pancreatic carcinoma, and myeloid leukemias (Barbacid, 1987, Ann. Rev. Biochem. 56:779). Approximately 20 to 30% of all human tumors can be attributed to the activation of the ras protooncogene.
- Ras genes constitute a multi-gene family that transform cells through the action of a 21 kDa protein termed ras p21 (also referred to herein as "ras").
- Ras functions as a G-regulatory protein, hydrolyzing GTP to GDP.
- ras In its inactive state, ras binds GDP.
- ras Upon activation of growth factor receptors, ras exchanges GDP for GTP and undergoes a conformational change.
- the wild-type ras couples the signals of activated growth factor receptors to downstream mitogenic effectors.
- the intrinsic GTP-ase activity of ras eventually returns the protein to its inactive GDP-bound state.
- a mutation in the ras gene results in a loss of regulatory function, resulting in constitutive transmission of growth stimulatory signals and oncogenic activation.
- ras For both normal and oncogenic functions, ras must be localized at the plasma membrane, a process that is dependent upon proper post-translational processing of the ras (Hancock, 1989, Cell 57:1167).
- a farnesyl group is attached to a cysteine residue at position 186 of the protein by reaction with farnesyl pyrophosphate .
- the carboxy-terminal three amino acids of the protein are cleaved by the action of a specific protease.
- the carboxylic acid terminus is converted to a methyl ester by alkylation with a methyl group.
- Post-translational modification of ras is mediated by an amino acid sequence motif frequently referred to as a "CAAX box."
- C represents Cysteine
- A represents an aliphatic amino acid
- X is another amino acid such as Methionine, Serine, or Glutamine .
- this motif serves as a signal sequence for farnesyl -protein transferase or geranylgeranyl-protein transferase, which catalyze the alkylation of the cysteine residue of the CAAX sequence.
- Farnesylation of ras is required for proteolytic processing, palmitoylation, and tight binding of the ras protein to cellular membranes.
- inhibitors of farnesyl-protein transferase have been shown to block the growth of ras-transformed cells in soft agar. Accordingly, inhibitors of farnesyl -protein transferase, and of ras activity in general, are thought to be useful anti-cancer therapeutics for many types of cancers (Gibbs et al . , 1984, Proc . Natl . Acad. Sci. USA 81:5704-5708; Jung et al . , 1994, Mol. Cell. Biol . 14:3707-3718; Predergast et al . , 1994, Mol. Cell.
- a pharmaceutical composition containing from about 40 mg to about 480 mg, preferably from about 175 mg to about 315 mg, more preferably from about 240 to about 280 mg of arglabin or a derivative thereof is provided in unit dosage form.
- the dose may be divided into 2-4 daily doses. Typical dosages of these pharmaceutical composition range from about 0.5 mg/kg to about 7 mg/kg. In extreme conditions, up to about 20 mg/kg may be administered.
- Lyophilized dimethylaminoarglabin and lyophilized pharmaceutically acceptable salts such as dimethylaminoarglabin hydrochloride are particularly useful as pharmaceutical compositions.
- the optimal concentration of arglabin or a derivative thereof in a pharmaceutically acceptable composition may vary, depending on a number of factors, including the preferred dosage of the compound to be administered, the chemical characteristics of the compounds employed, the formulation of the compound excipients and the route of administration.
- the optimal dosage of a pharmaceutical composition to be administered may also depend on such variables as the type and extent of cancer metastases, the overall health status of the particular patient and the relative biological efficacy of the compound selected.
- These compositions may be used for the treatment of cancer, especially lung, liver and ovarian cancer, although other cancers such as breast, rectal, colon, stomach, pancreatic or esophageal cancer are also beneficially treated with the compositions.
- hematopoietic cancers such as leukemias and lymphomas may also be beneficially treated.
- Compounds of the invention may be formulated into pharmaceutical compositions by admixture with pharmaceutically acceptable non-toxic excipients or carriers.
- Such compounds and compositions may be prepared for parenteral administration, particularly in the form of liquid solutions or suspensions in aqueous physiological buffer solutions; for oral administration, particularly in the form of tablets or capsules; or for intranasal administration, particularly in the form of powders, nasal drops, or aerosols.
- Compositions for other routes of administration may be prepared as desired using standard methods.
- a compound of the invention may be conveniently administered in unit dosage form, and may be prepared by any of the methods well known in the pharmaceutical art, for example, as described in Remington's Pharmaceutical Sciences (Mack Pub. Co., Easton, PA, 1980) .
- Formulations for parenteral administration may contain as common excipients sterile water or saline, polyalkylene glycols such as polyethylene glycol, oils of vegetable origin, hydrogenated naphtalenes, and the like.
- polyalkylene glycols such as polyethylene glycol, oils of vegetable origin, hydrogenated naphtalenes, and the like.
- biocompatible, biodegradable lactide polymer, lactide/glycolide copolymer, or polyoxethylene- polyoxypropylene copolymers are examples of excipients for controlling the release of a compound of the invention in vivo .
- Other suitable parenteral delivery systems include ethylene-vinyl acetate copolymer particles, osmotic pumps, implantable infusion systems, and liposomes.
- Formulations for inhalation administration may contain excipients such as lactose, if desired.
- Inhalation formulations may be aqueous solutions containing, for example, polyoxyethylene-9- lauryl ether, glycocholate and deoxycholate, or they may be oily solutions for administration in the form of nasal drops. If desired, the compounds can be formulated as a gel to be applied intranasally. Formulations for parenteral administration may also include glycocholate for buccal administration.
- the invention also relates to an article of manufacturing containing packaging material and arglabin or a derivative thereof contained within the packaging material .
- Arglabin or derivatives thereof are therapeutically effective for suppressing tumor growth in a human.
- the packaging material contains a label or package insert indicating that arglabin or a derivative thereof may be used for suppressing tumor growth in a human.
- Dimethylaminoarglabin and pharmaceutically acceptable salts thereof are arglabin derivatives that are particularly useful in the article of manufacture.
- the invention relates to compositions and kits comprising a first chemotherapeutic agent including arglabin or a derivative thereof and a second chemotherapeutic agent.
- the second chemotherapeutic agent is not arglabin or a derivative thereof.
- These compositions are effective to suppress tumor growth in a human.
- Dimethylaminoarglabin or a pharmaceutically acceptable salt thereof is a particularly useful derivative of arglabin.
- chemotherapeutic agents including alkylating agents, antimetabolites, vinca alkaloids, antibiotics or platinum coordination complexes may be used in the composition.
- alkylating agents such as the nitrogen mustards cyclophosphamide and sarcolysin may be used, although other alkylating agents such as methylnitrosourea are also appropriate.
- Antimetabolites such as the folic acid analog methotrexate or pyrimidine analogs such as fluorouracil or 5-fluorouracil may be used, as well as vinca alkaloids such as vinblastine or vincristine.
- An antibiotic such as rubidomycin can be an appropriate chemotherapeutic agent, as well as platinum coordination complexes such as cisplatin.
- Multiple chemotherapeutic agents may be combined with arglabin or a derivative thereof.
- vincristine and cyclophosphamide or vincristine and vinblastine may be combined with arglabin or a derivative thereof.
- the invention also relates to a method of suppressing tumor growth in a human patient by administering to the patient an amount of a composition including a first chemotherapeutic agent including arglabin or a derivative thereof and a second chemotherapeutic agent effective to suppress tumor growth in the human.
- the second chemotherapeutic agent is not arglabin or a derivative thereof.
- These compositions provide an enhanced antitumor effect and may also prevent development of metastases . In particular, these compositions are useful for overcoming tumors that are drug-resistant.
- the agents may be administered separately or as a cocktail. Toxicity may be reduced by administering arglabin or a derivative thereof several hours prior to administering the chemotherapy agent.
- the compositions may be administered by any route.
- the invention also relates to a method for reducing the immundepressive effect of a chemotherapy agent in a human patient by administering to the patient an amount of arglabin or a derivative thereof effective to augment the immune system of the patient upon treatment of the patient with the chemotherapy agent.
- the immune system may be augmented, for example, by increasing the total number of leukocytes, T-lymphocytes, B-lymphocytes, or immunoglobulins .
- Example 1 Isolation of Arglabin -
- the smooth wormwood Artemisia glabella Kar. et Kir. is a perennial plant that is widespread on the Ukraine dry steppe hills.
- the solvent vessel has a filter, distillator with an evaporator and condenser, and a buffer capacity.
- the drying agent vessel consists of a dryer, cyclone, cooler, ventilator and heater.
- the cooling water vessel includes a saltpan with ventilation.
- the extraction device also has a deodorizer with ventilator, a waste tank and an extract collector. Approximately 7.7 kg of dry material from Artemisia glabella Kar. et Kir. was placed in the extraction device and continuously mixed with solvent as the material was moved through the extractor column. The solvent moves in the opposite direction of the dry plant material and gradually becomes saturated with extracted substances.
- the saturated solvent was discharged, it was first filtered to remove plant material particles, then evaporated. The filtered plant particles were recirculated through the extractor for re-extraction. Vapors from the evaporation were sent to the condenser. From the condenser, pure solvent was recovered and recirculated to the extraction device. Condensation surfaces in the condenser were cooled with water pumped from the salt pan where the water was previously cooled with exterior air blown in the ventilator. Due to air- vaporized cooling in the salt pan, the water may be cooled down to temperatures considerably lower than the ambient temperature .
- the extracted substances refined from the solvent are in the form of a tar. During this process, approximately 7% of the plant material (539 grams) was recovered.
- the tar was further refined by addition of two volumes (approximately 1.08 L) of 60°C ethanol with continuous stirring to dissolve the tar. Distilled water, heated to approximately 70°C, was added in a ratio of about 2:1 alcohol to water. The tar-alcohol -water solution was thoroughly stirred for 30 minutes, then left at room temperature for approximately 24 hours or until a precipitate was formed. The water alcohol solution was filtered through a ceramic filter under vacuum. The procedure was repeated with any precipitate remaining after filtration.
- the filtrate was rotary evaporated and the alcohol was vacuum distilled in the form of an azeotropic mixture with water containing 68-70% alcohol. After distillation of the alcohol, the water solution yielded approximately 286 grams of refined tar.
- the refined tar was separated into individual components over a KCK silicagel column, with pressure, using benzene as the eluant .
- Benzene fractions were collected and analyzed for arglabin using thin-layer chromatography (TLC) (silufol, benzene-ethanol, 9:1).
- Arglabin-containing fractions were distilled to remove benzene. Arglabin at this stage has a yellow color.
- Approximately 33 g of arglabin was produced, with a yield of about 11.7%.
- Arglabin was recrystallized by dissolving in hexane in a 1:10 ratio of product to hexane (w/v) and heating. After arglabin was in solution, the product was vacuum filtered. Crystals of arglabin were isolated from the filtrate at room temperature. Approximately 21 g of arglabin was recovered from this step.
- Arglabin has a structure of 1 (R) , 10 (S) -epoxy-5 (S) , 6 (S) , 7 (S) -guaia- 3(4), 11 (13) -diene-6, 12-olide. The stereochemistry of arglabin was determined through x-ray analysis.
- Example 2 Arglabin Derivatives - To assist the reader, the names of the various compounds set out below are followed with numerals to facilitate identification with the compounds depicted in the Figures.
- dichlorohydrines 3 and 4 Treatment of epoxyarglabin 2 with an ether- acetone HCl solution produced dichlorohydrines 3 and 4 ( Figure 1) . An opening of both epoxygroups with a yield of 60% and 95% regioselectivity was observed. Dichlorohydrines 3 and 4 were diluted with water, washed with NaHC0 3 , and purified by silica gel column chromatography using Et 2 0-iPr 2 0 in a 1:1 ratio. IR and NMR spectral data were used to confirm the structure .
- the 1,10-epimer of arglabin, epiarglabin 13 was synthesized by adding approximately 0.1 ml of P0C1 3 to a cooled solution (approximately 0°C) of 120 mg of diol 10 in pyridine. ( Figure 2) After stirring for 24 hours at approximately -5°C, the reaction was worked up by extraction with ethyl ether. After washing with 5% HCl and water, the residue was crystallized from petroleum ethyl ether to give 40 mg of 1, 10-epiarglabin 13.
- Dimethylaminoarglabin 14b was purified by first dissolving in 5 volumes (w/v) of chloroform then mixing with about 3 volumes (w/w) of KCK silica gel. After evaporation of the solvent, the dry material was chromatographically separated on a KCK silica gel column made with a 1:22 ratio of adduct to sorbent . The column was eluted by a mixture of petroleum diether and sulfuric ether (1:1, 1:2). Fractions of approximately 14-17 mis were collected and monitored with TLC . Dimethylaminoarglabin 14b was recrystallized from the fraction with chloroform and ether (1:1) .
- Dimethylaminoarglabin hydrochloride 14d was produced by dissolving dimethylaminoarglabin 14b with 0.22 L of alcohol and heating to 40°C. After vacuum filtration, hydrogen chloride gas was produced by addition of 0.2 kg of sodium chloride and drops of concentrated sulfuric acid. The reaction was monitored by TLC. When the reaction was complete, the mixture was heated to 52 °C and the ethanol was vacuum distilled. Approximately 0.9 L of ethylacetate was added to the remaining tar with intensive stirring. The resulting precipitate yielded approximately 21 g of dimethylaminoarglabin hydrochloride 14d.
- Example 3 Lyophilization - The water solution of dimethylaminoarglabin hydrochloride was filtered through a cotton-gauze plug or 8 layers of gauze, and a sterile Millipor filter to a sterile glass jar. The solution was vacuum pumped out of the jar into a measuring buret and aliquoted into 2 ml vials or ampules. The filled vials or ampules were maintained at -40°C on sterile shelves for 24 hours prior to drying in a KC-30 lyophilizer or a LS-45 lyophilizer. After this tempering period, the drying process was started.
- the temperature was maintained at -40 °C for 2 hours, then was gradually increased to approximately 50°C (plus or minus about 5°C) .
- the transition to approximately 50°C occurred over about 12-13 hours of drying.
- the final temperature did not exceed +60 °C.
- the total duration of drying time was 24 hours.
- the vials with dry compound were immediately covered with caps and rolled. Ampules were soldered. Each vial or ampule contained about 0.04 g of the preparation.
- Vials or ampules that were not sterile filtered were sterilized by autoclaving for 20 minutes at 120°C, with pressure of 1.2 Atm.
- the prepared dimethylaminoarglabin hydrochloride water solution was filtered through a cotton-gauze plug or 8 layers of gauze. Approximately 200 ml of the solution were poured into 500ml bottles, covered with cotton-gauze plugs and wrapped with oilpaper. The filled bottles were sterilized by autoclaving for 30 minutes at 120°C with 1.2 Atm of pressure. The sterile solution was cooled to room temperature. Using sterile technique, 2 ml of the solution was poured into sterile 10ml vials. The vials were then lyophilized as described above. After lyophilization each vial contained about 0.04g of the compound.
- the lyophilized material had a white-straw color and a bitter taste. Authenticity of the preparation was verified by determining its melting point and recording IR- , mass-, and NMR-spectra. The quality of the preparation was controlled by diluting 1 mg of the preparation with 0.2 ml of water. Addition of one drop of a saturated vanillin solution in concentrated sulfuric acid turned the mixture a violet color, indicating the presence of terpenes . Lyophilized material may be stored for three years .
- Glabellin 17 was also isolated from Artemisia glabella Kar. et Kir. The yield of the compound from dry raw material was approximately 0.016%. The structure of glabella was determined through IR- , UN-, NMR, C13 NMR- , mass-spectra and chemical transitions.
- Glabellin Characteristics melting point 130-131°C (petrol-diether) ; [ot] 20 D +90.9° (SO, 17, chloroform) .
- 3-keto-eudesm-l (2) , 4(5), 11 (13) -trien-6, 12- olid(l) 18 was prepared by selective dehydration of ⁇ - sautonine with a yield of 45% and may be produced from more than 20 species of wormwood.
- the structure was determined by IR- , UV- and NMR-spectra.
- Epoxy estafiaton 20 was produced by isomerizing an available terpene lactone such as estafiatine through isomerization with etherate of trifluoride boron then epoxidizing with re-chlorbenzoil acid.
- the 3-keto- 10c(14) -epoxy-l,5,7 ⁇ (H) 4,6/3(H) -guai-11 (13) -en-6, 12-olid structure was determined by IR- , NMR- and mass-spectra.
- Gaigranin 21 was produced by extraction of the aerial part of Gaillardio grandiflora with chloroform then chromatographically separating on a silica gel column. The structure of gaigranin 21 was confirmed by IR-, UV-, NMR and Cesy-spectra.
- Example 5 Jn-vitro Activity of Arglabin and Derivatives -Viability of Cells - Transformed cells and primary cultures of normal cells were incubated with varying concentrations of dimethylaminoarglabin hydrochloride to determine its effect on the viability of the cells.
- Mouse mastocytoma P-815) , myeloma (Z-P3x63Ag8.653 and Pai) and human erthyroleukemia (K-562) cell lines were used.
- Primary cultures of normal mouse hepatocytes were isolated from mouse liver using collagenase.
- Mouse splenocytes were isolated using a glass homogenizer.
- Marrow cells were obtained by washing the bone marrow. See, for example, Shears, S.B. and Kirk, CJ. (1984), Biochem. J. 219:375-382.
- Cells were cultured in RPMI-1640 medium supplemented with 10% fetal calf serum, 100 mM L- glutamine and 50 ⁇ g/ml gentamycin at 37°C under 5% C0 2 . Cells were seeded into 24 -well plates at a density of 50,000 cells/well and grown until near confluency, approximately 2 days, then transferred to 96-well plates at the same density. Transformed cell lines were incubated for 18 hours with dimethylaminoarglabin hydrochloride, in concentrations ranging from 1.5 ⁇ g/ml to 100 ⁇ g/ml. Viability of the cells was determined by trypan blue exclusion.
- the proliferation of the transformed cells was assessed by incubating 3 H-labeled thymidine in the media for 18 hours. At the end of the specified time period, the proliferation was measured by counting the amount of 3 H-thymidine incorporated. Thymidine incorporation provides a quantitative measure of the rate of DNA synthesis, which is typically directly proportional to the rate of cell division.
- Figure 6 shows that proliferation of X-653 and P-815 cells was effectively blocked at concentrations of 6 ⁇ g/ml and 12 ⁇ g/ml, respectively.
- Primary cultures of normal cells were incubated for 18 hours with a concentration of dimethylaminoarglabin hydrochloride ranging from 10 ⁇ g/ml to 2560 ⁇ g/ml. Viability was measured by trypan blue exclusion.
- Figure 7 shows that an increase in the concentration of dimethylaminoarglabin hydrochloride reduced the viability of the normal cells, but a much higher concentration was necessary to kill the normal cells, as compared to the transformed cells.
- concentration of 320 ⁇ g/ml the number of viable splenocytes was reduced by 50% in comparison to the control.
- concentrations of 640 ⁇ g/ml and 1280 ⁇ g/ml 40% and 10%, respectively, of the splenocytes were still viable. At these same concentrations, approximately 50% and 25% of hepatocytes were still viable. Marrow cells were more sensitive to dimethylaminoarglabin hydrochloride.
- Protein Prenylation - Mouse myeloma Pai cells were cultured in the presence of 60 ⁇ M dimethylaminoarglabin hydrochloride. The cells were collected by centrifugation at 600 x g for 10 minutes and then washed twice in PBS. Control cells were grown in the absence of drug. The cells were solubilized in lysis buffer (50mM Tris, pH 7.4, 25 mM EDTA, 0.05% Tween, 0.01 M NaCl) for 30 minutes on ice.
- lysis buffer 50mM Tris, pH 7.4, 25 mM EDTA, 0.05% Tween, 0.01 M NaCl
- Lysates were made by homogenization for 5 minutes at 4°C and precipitated by centrifugation at 12,000 x g for 10 minutes. The supernatant was collected. Proteins were precipitated with trichloroacetic acid and then successively washed with ethanol and ethyl ether. A selective naphthol cleavage of the bond between isoprenoides and proteins was performed as described by Epstein, W.W et al . , (1991) Proc. Natl. Acad. Sci . USA 88:9668-9670. In general, 5 mg of a potassium naphthoxide and naphthol 4:1 mixture was added to approximately 10 mg of precipitated protein.
- the molar ratio of geranylgeranyl to farnesylcysteine was 6.
- the influence of dimethylaminoarglabin hydrochloride on cellular prenylation is shown in Figure 8B.
- the peak corresponding to the farnesylcysteine derivative does not appear on the chromatogram, while the geranylgeranyl peak appeared as in the control . This indicates that dimethyl -aminoarglabin hydrochloride can prevent farnesylation of proteins without significant effects on geranylgeranylation.
- the compounds in this family have low toxicity and are tolerated at dosages exceeding the therapeutic dosage.
- Conventional toxicology methods were used to determine the LD 50 for an intraperitoneal injection of a 2% solution of dimethylaminoarglabin hydrochloride in dimethylsulfoxide (DMSO) in mice (weight 20-22 g) and rats (120-130 g) .
- the LD 50 was 190-220 mg/kg in mice and 280-310 mg/kg in rats.
- An autopsy of the animals revealed plethoricy of internal organs, vasodilatation of the mesentery and intestines.
- Tolerant single doses in rats and rabbits did not disturb the function of the liver, kidneys, cardiovascular system, respiration or peripheral nervous system. Blood pressure was maintained. In addition, no pyrogenic, allergenic, teratogenic or embryo toxic effects were observed in animals.
- MTD tolerable doses
- arglabin and derivatives were determined by daily intraperitoneal administration to rats, guinea pigs or mice and daily intravenous administration to rabbits over a period of five to 20 days.
- the MTD ranged from about 20 mg/kg to about 50 mg/kg for all compounds tested.
- the maximum dosage of dimethylaminoarglabin hydrochloride in a solution of DMSO ranged from 20 mg/kg in rabbits, 30 mg/kg in mice, 45 mg/kg in guinea pigs to 50 mg/kg in rats.
- Dimethylaminoarglabin and dimethylaminoarglabin hydrochloride were effective against a wide range of tumors .
- One advantage of dimethylaminoarglabin hydrochloride is that it is soluble in water.
- this combination overcame the cross-resistance of sarcoma-45 to methotrexate, sarcoma-45 to 5-Fluorouracil , and Pliss' lymphosarcoma to rubidomycin. No animals deaths were observed with this treatment.
- the combination of dimethylaminoarglabin hydrochloride and methylnitrosourea was administered at intervals of 2 , 4 or 24 hours between the two drugs. It was determined that it was optimal to administer dimethylaminoarglabin hydrochloride two hours prior to administration of methylnitrosourea.
- the cross resistance of sarcoma-45 to prospidin and sarcoma-45 to 5-fluorouracil, Pliss' lymphosarcoma to rubidomycin and Pliss' lymphosarcoma to prospidin was overcome with the combination of dimethylaminoarglabin hydrochloride and methylnitrosourea. Approximately 60% of the tumors disappeared in the rats without adverse drug reactions.
- Dimethylaminoarglabin hydrochloride moderately increased the duration of life in the animals with non- resistant and drug resistant tumors.
- the combination of dimethylaminoarglabin hydrochloride and other antitumor drugs further prolonged the duration of life.
- the combination of dimethylaminoarglabin hydrochloride and vincristine prolonged life 114% in animals with methylnitrosourea resistant tumors.
- a good therapeutic effect was seen from the triple combination of dimethylaminoarglabin hydrochloride, vincristine and cyclophosphamide at half of MTD as compared with the double combination of dimethylaminoarglabin hydrochloride and vincristine or dimethylaminoarglabin hydrochloride and cyclophosphamide.
- the triple combination prolonged duration of life by 209%.
- the quadruple combination of dimethylaminoarglabin hydrochloride, vincristine, cyclophosphamide and cisplatin was less effective than the triple combination. This may be due to increased toxicity of the antitumor drugs.
- the effect of dimethylaminoarglabin hydrochloride alone and in combination with other drugs was studied in the model of drug resistant metastasis of Pliss' lymphosarcoma.
- the metastases in the inguinal lymphoid nodes were the most sensitive among the initial and drug resistant nodes. They did not develop in 10% of the cases.
- the duration of life for dimethylaminoarglabin hydrochloride alone was 128% in comparison with the control group.
- the combination of dimethylaminoarglabin hydrochloride and vincristine caused inhibition of tumor growth, with tumor dissolution, in 30% of the rats. Duration of life was increased 174% with the absence of any new metastases in the inguinal lymphoid nodes .
- dimethylaminoarglabin hydrochloride and sarcolysin were used for the treatment of sarcoma 45 to investigate the disturbance of DNA synthesis.
- Beneficial results were observed with sarcolysin and with the combination of sarcolysin and dimethylaminoarglabin hydrochloride in the case of the non-drug resistant sarcoma 45.
- dimethylaminoarglabin hydrochloride alone was very effective (DNA inhibition index 99%) .
- dimethylaminoarglabin hydrochloride increased immunological indices, particularly if dimethylaminoarglabin hydrochloride was administered two hours before the cytostatic drugs .
- dimethylaminoarglabin hydrochloride softened the immunodepressive effect of cytostatics and normalized the immune balance of the body.
- dimethylaminoarglabin hydrochloride decreased cytotoxicity and increased the efficacy against drug resistant tumors alone and in combination.
- mice The effect of dimethylaminoarglabin hydrochloride was determined in intact and immunodepressed mice. Mice were immunodepressed by administration of 200 mg/kg of cyclophosphamide. Injection of cyclophosphamide resulted in considerable leukopenia related primarily to lymphopenia. The humoral immunity of the animals was considerably suppressed, as was cell -mediated immunity although to a lesser extent. Two dosages of a 2% dimethylaminoarglabin hydrochloride solution, 50 and 100 g/kg, were injected IP into white mongrel mice. The Hemagglutination test and delayed-type hypersensitivity reaction were determined before and after administration of drug.
- T-lymphocytes Daily IP administration of 10 and 20 mg/kg dimethylaminoarglabin hydrochloride for 10 days to intact mice resulted in a dramatic change in T-lymphocytes, B- lymphocytes and natural killer cells.
- the overall leukocyte count was increased through an increase in natural killer cells and T-lymphocytes while B- lymphocytes remained stable. It was found that the increase in T-lymphocyte number was a result of an increase in the level of the T-helper subpopulation. The level of the T-suppressor subpopulation was not altered.
- a higher dosage (20 mg/kg) overall leukocyte number was not altered, even though B-lymphocyte number decreased and T-lymphocyte decreased to a lesser extent. Natural killer cell number was increased.
- dimethylaminoarglabin hydrochloride increased T and B- lymphocytes and natural killer cell levels.
- the increase in T-lymphocytes was accompanied by an increase in the level of the T-helper subpopulation of T-lymphocytes .
- Higher dosages of dimethylaminoarglabin hydrochloride (20mg/kg) decreased T and B-lymphocyte number, but increased certain other cell populations, such as natural killer cells.
- lyophilized-dimethylaminoarglabin hydrochloride stimulated delayed type hypersensitivity, but reduced all other studied indices in intact rats. In rats with Pliss Lymphosarcoma, the immune system was stimulated, as all studied indices increased.
- An advantage of lyophilized- dimethylaminoarglabin hydrochloride is that it ameliorates the immunodepressi e effect of known cytostatics such as 5-fluorouracil and sarcolysin.
- Dimethylaminoarglabin hydrochloride was excreted fairly slowly. The biological half-life was about 46.8 hours in rats, with the average time of retention about 67 hours. Renal excretion proceeded slowly. The kidney concentration was maximal after three hours. By 24 hours, the kidneys had the highest concentration, 56.6 ⁇ g/g. Total clearance was 0.05 ml/minute at a low transportation rate of the preparation from peripheral tissues into the blood.
- dimethylaminoarglabin hydrochloride was administered intraperitoneally. Intrapleural administration was used for patients with pleurisy. Patients were given a very small dose and monitored for any signs of allergic reaction before proceeding. Initially, 80 mg of the compound was given per day as a single dose, then gradually increased to a maximum level. After 30-35 days, the dose was increased to 480 mg per day. At this high dose, patients complained of nausea and vomiting. It was estimated that the daily dose should be about 240- 280 mg for typical cases. Total dose over the course of treatment was typically five to six grams of dimethylaminoarglabin hydrochloride, but was as high as 20 grams.
- Tx - tumor cannot be adequately assessed, TO - no evidenc
- N (Regional lymph node) , Nx - regional lymph nodes cannot be assessed clinically, NO - no evidence of regional node metastasis, Nl, N2, N3 - increasing involvement of region lymph nodes
- M subclasses disant metastasis
- Mx - not assessed MO - no distant metastasis
- MO - no distant metastasis MO - no distant metastasis present, specify site(s)
- Dimethylaminoarglabin hydrochloride had little toxicity and did not suppress hematopoiesis . During the trial, no negative responses of the gastrointestinal tract or hair follicles were registered.
- Immunologic status of the patients was evaluated using standard methods of rozette-formation and phagocytosis. These indices were studied prior to treatment, during treatment and after treatment. Blood samples were taken from a finger. Analysis of the average immunological 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 adhesiveness decreased from 42% to 28.5%. Undifferentiated lymphocytes increased from 21.5% to 42.2%. A general change in the ratio of T-helper lymphocytes to T- suppressor lymphocytes was due to an increase of T- suppressor lymphocytes.
- the number of B-lymphocytes and phagocytic activity remained stable. Total number of leukocytes increased up to 9.4xl0 9 /L and the total number of lymphocytes increased as well. The levels of all types of immunoglobulins increased.
- indices returned to normal. In some patients, indices returned to normal by day 14. In patients that were analyzed 30 days after treatment, a significant increase in the number of T- lymphocytes and adhesiveness of neutrophils was observed.
- dimethylaminoarglabin hydrochloride was given to 72 patients (61.1% male and 38.9% female) with stage IV cancer from different localizations. Among the patients, 25% had carcinoma of the stomach, 16.7% had liver cancer, 18.1% had lung cancer and the remaining 40.2% had esophageal, breast, ovarian, pancreatic, brain or lymphosarcoma. Patients with poorer states had metastases to the liver (25%) , retroperitoneal lymph nodes (25), ascites (22.2%) and exudative pleuritis (11.1%). Some of these patients had been previously treated with dimethylaminoarglabin hydrochloride in the first clinical trial. Results from the second clinical trial are summarized in Table VII.
- dimethylaminoarglabin hydrochloride as an anti-tumor cytostatic in solid tumors has a number of advantages.
- the preparation has no side effects, it does not suppress hematopoiesis, it normalizes the functional condition of immune system, and has no allergenic effect.
- As a cytostatic it is particularly efficient for primary cancer of the liver and other solid tumors complicated with polyserositis . Partial regression of tumor was observed in 61.1% cases; stabilization of process - in 31.9% cases and recurrence was observed in only 7.0%. 88.9% of patients (64 of 72) responded to therapy: no response was observed in 11.1% (8 of 72).
- Patient M age 55, case number 305, entered the hospital with multiple nodes on the skin of thorax and abdomen, ulcer on the place of extripated breast, and induration on the right breast .
- a radical mastectomy had been performed at Sakhalinsk Oncology Center due to breast cancer.
- she received 6 courses of polychemotherapy with cyclophosphamide and methotrexate.
- Symptomatic therapy was recommended because of recurrence of the process.
- the abdomen and thorax skin had multiple metastatic nodes with sizes ranging from 0.5 to 1 cm.
- an ulcerous surface approximately 10x12 cm, was present.
- the right breast was deformed because of infiltrative metastases. Edema was present in the lower extremities.
- a blood analysis before treatment revealed the following parameters: Hb-89, ESR-6 mm/hy, L-3.3, Er-3.8 ml, juv.ne-4, seg. ne-78, mon-1.
- the patient received 5 courses of dimethylaminoarglabin hydrochloride treatment at a total dose from 6.0 to 7.3 grams.
- the ulcer was epithelized, the metastases nodes were resolved, and infiltration of the right breast decreased 50%.
- Ultrasonic tomography (UST) of the liver indicated that the liver was enlarged and occupied the whole abdominal cavity.
- the structure was dissimilar because of the foci of dissimilar structure with hydrophilic rim up to 5-6 cm, indicating a liver tumor.
- Blood analysis before treatment revealed the following parameters: Hb-84, ESR-4 mm/h, L-10.9, Er-3.3 ml, eos-1, juv, ne-55, stab ne-45, seg, ne-14, lym-30, mon-5.
- Paracentesis of the liver was performed under the control on UST. Bare nuclei of tumor cells were observed against a background of hepatic cells with degenerative changes. The patient was diagnosed with embryonal liver cancer.
- the total dose for the course of treatment was 2040 mg.
- the abdomen became symmetric and smaller due to the decrease in liver size.
- a UST indicated that the liver projected from under the coastal arch along midclavicular line on 4 cm, the outlines are even, the structure is dissimilar because of the foci of dissimilar structure with hydrophylic rim and foci of high echogenity 2.0-2.5 cm in diameter.
- the indices of immune system were determined using rozette forming and phagocytosis methods. 57 patients that had received dimethylaminoarglabin hydrochloride were examined. Eleven indices of cellular and humoral immunity were measured from each patient in order to evaluate immune status. The following indices were determined on 0.05 ml of peripheral blood: absolute and relative amount of T- and B-lymphocytes, amount of non- differentiated "zero" cells, adhesion and phagocyte activity of neutrophils, hemogram, level of serum immunoglobulins . Indices were determined before treatment, on days 2, 5 and 14 of treatment, and after treatment. Table X summarizes the results before and after treatment .
- the total number of lymphocytes in the peripheral blood was elevated. As tumors can cause both quantitative and qualitative changes in blood cells, these parameters were checked after dimethylaminoarglabin hydrochloride treatment. No shift in qualitative (morphological) composition of blood cells was identified, although some quantitative changes such as a decreased number of neutrophils and an increased number of lymphocytes was observed. This suggests a reduction lymphotoxic effects caused by the tumor.
- the immunologic indices correlated with the clinical findings in most cases .
- Mean values of immunological indices were evaluated with regression analysis. Functional conditions of the immune system were estimated using integral indices such as a mean intensity (correlation) expressed in relative units. That the index-intensity of the immune system increased during treatment indicates that the immune system responded to treatment. Correlation analysis of these data indicate that during treatment, the total amount of true bound parameters increased (the number of bonds with r> 0.7 increased, and the number of negative bonds was reduced) . The number of interrelations between the elements of immunity increased, namely between the lymphocyte and neutrophil elements.
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US93422897A | 1997-09-19 | 1997-09-19 | |
US934229 | 1997-09-19 | ||
US934471 | 1997-09-19 | ||
US934228 | 1997-09-19 | ||
US08/934,471 US6693127B1 (en) | 1997-07-03 | 1997-09-19 | Pharmaceutical compositions of arglabin and arglabin derivatives |
US08/934,229 US5902809A (en) | 1997-07-03 | 1997-09-19 | Arglabin compounds and therapeutic uses thereof |
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US6051565A (en) * | 1997-04-26 | 2000-04-18 | International Phytochemistry Research Labs, Ltd. | Farnesyl-protein transferase inhibitors |
GB9809889D0 (en) * | 1998-05-11 | 1998-07-08 | Univ Strathclyde | Prevention of restenosis |
WO1999064003A1 (en) * | 1998-06-09 | 1999-12-16 | Tovarischestvo S Ogranichennoi Otvetstvennostju 'tabifarm' | Method of treatment of patients with initial liver cancer |
CA2356438A1 (en) * | 2001-09-05 | 2003-03-05 | Andre Pichette | Use of terpenes and derivatives as potentiators of antitumor agents in the treatment of cancers |
DK1643984T3 (en) * | 2003-07-11 | 2011-10-24 | Univ Kentucky Res Found | Use of parthenolide derivatives as antileukemic and cytotoxic agents |
US7678904B2 (en) | 2003-07-11 | 2010-03-16 | University Of Kentucky | Use of parthenolide derivatives as antileukemic and cytotoxic agents |
CN102372723B (en) * | 2010-08-19 | 2014-03-26 | 石药集团中奇制药技术(石家庄)有限公司 | Method for extracting arglabin from artemisia myriantha |
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Title |
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BOTTEX-GAUTHIER C ET AL: "IN VITRO BIOLOGICAL ACTIVITIES OF ARGLABIN, A SESQUITERPENE LACTONE FROM THE CHINESE HERB Artemisi myriantha WALL. (ASTERACEAE)" BIOTECHNOLOGY THERAPEUTICS, MARCEL DEKKER, NEW YORK, NY, US, vol. 4, no. 1/2, 1993, pages 77-98, XP002114696 ISSN: 0898-2848 * |
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