WO2000061153A1 - Compositions medicinales et leur procede de preparation - Google Patents

Compositions medicinales et leur procede de preparation Download PDF

Info

Publication number
WO2000061153A1
WO2000061153A1 PCT/AU2000/000300 AU0000300W WO0061153A1 WO 2000061153 A1 WO2000061153 A1 WO 2000061153A1 AU 0000300 W AU0000300 W AU 0000300W WO 0061153 A1 WO0061153 A1 WO 0061153A1
Authority
WO
WIPO (PCT)
Prior art keywords
composition
glycoalkaloid
group
bec
preparation
Prior art date
Application number
PCT/AU2000/000300
Other languages
English (en)
Inventor
Bill E. Cham
Original Assignee
Cura Nominees Pty. Ltd.
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Cura Nominees Pty. Ltd. filed Critical Cura Nominees Pty. Ltd.
Priority to AU35454/00A priority Critical patent/AU779512B2/en
Priority to EP00913972A priority patent/EP1181022A4/fr
Priority to CA002369272A priority patent/CA2369272A1/fr
Publication of WO2000061153A1 publication Critical patent/WO2000061153A1/fr

Links

Classifications

    • 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
    • A61K31/702—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
    • 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
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
    • 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
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
    • 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
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/12—Keratolytics, e.g. wart or anti-corn preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention relates to medicinal compositions and in particular therapeutic compositions comprising glycoalkaloids .
  • Such compositions may be used in the treatment, control and diagnosis of cancers and tumors in mammals, contraception and termination of pregnancy.
  • the present invention is particularly directed towards a composition comprising a mixture of solasodine glycosides.
  • the present invention is also directed towards a method of preparing a medicinal composition and a method of treatment, control or diagnosis of cancers and tumors in mammals.
  • Glycoalkaloids are steroidal alkaloids which have a sugar moiety bound to the alkaloid moiety.
  • the sugar moiety can be a monosaccharide, disaccharide, oligosaccharide or polysaccharide .
  • Certain glycoalkaloids derived from plants have been observed to have anti-cancer properties.
  • glycoalkaloids extracted from the Solanum genus are glycoalkaloids extracted from the Solanum genus .
  • Glycoalkaloids from the species Solanum Sodomaeum L have been shown to be active against cancer in animals and skin tumors in humans.
  • the glycoalkaloids extracted from the fruit of Solanum Sodomaeum L are glycoalkaloids extracted from the fruit of Solanum Sodomaeum L .
  • solasonine [ (22R, 25R) - spiro-5-en-3 ⁇ -yl- ⁇ -L- rhamnopyranosyl- (l->2gal) -O- ⁇ -D-glucopyranosyl- (l->3gal) - ⁇ -D-galactopyranose] (33%) , solamargine' (22R, 25R) -spiro-5- en-3 ⁇ -yl- ⁇ -L-rhamnopyranosyl- (l->2glu) -O- ⁇ -L- rhamnopyranosyl- (l->4glu) - ⁇ -D-gluco-pyranose] (33%) , and their corresponding di- and monoglycosides (34%) .
  • glycosides contain the same aglycone, solasodine.
  • This mixture of glycosides which includes solasonine and solamargine is commonly referred to as BEC.
  • BEC The structures of solasonine and solamargine are shown below:
  • BEC anti-cancer properties of BEC has been studied in vivo with mice inoculated with murine sarcoma 180 and in cell culture studies. BEC was observed to selectively destroy tumor cells relative to normal cells. The efficacy and specificity of BEC was also observed to be dependent upon the type of tumor. These observations were attributed to the presence of endogenous endocytic lectins or EEL's present on the membranes of those cells observed to be susceptible to BEC. EEL's are endogenous receptors which have been reported to be expressed during human embryogenesis and carcinogenesis. Interaction of the EEL with a molecule or ligand for which it is a receptor results in internalization of the EEL and bound ligand.
  • the tumor cells susceptible to BEC have EEL receptors specific for the glycoside portion of the glycoalkaloids in BEC. These EEL's selectively recognize and bind the sugar moiety of the glycoalkaloid. The glycoalkaloid is subsequently internalized and the result is destruction of the cell. The mechanism of cell destruction is believed to be by cell lysis. That there is an EEL specific for the glycoside moiety of the glycoalkaloid is supported by a number of observations. First, the aglycone, solasodine, when administered at levels at which BEC is effective is ineffective against tumor cells. The sugar portion of the glycoalkaloid on its own is also ineffective.
  • mice with murine sarcoma 180 Untreated mice died in 2-3 weeks.
  • Four doses of 8mg/kg BEC given on consecutive days resulted in survival of virtually all animals .
  • Five mg rhamnose/kg decreased the survival to 75%, lOmg rhamnose/kg decreased the survival to 50% and 15mg rhamnose/kg decreased the survival to 42%.
  • Similar concentrations of rhamnose were observed to have no effect on S180 activity in the absence of BEC.
  • ip intraperitoneal
  • the quantal effective levels (ED 50 ) of BEC for single administrations were similar to the lethal levels (LD 50 ) for multiple administrations of lOmg/kg at 14 daily ip doses .
  • BEC has also been observed to be effective for melanoma and ovarian tumor cells grown in cell culture.
  • the therapeutic index (LD 50 /ED 50 ) for these cell culture trials was about 3.
  • BEC a disadvantage of BEC is the toxicity of the preparation when administered at the very high levels required to successfully treat internal cancers. It would therefore be desirable to obtain a glycoalkaloid composition which is more effective than BEC for the treatment of cancers .
  • mice with advanced cancer activity could tolerate up to three times the LD 100 of BEC. This could be explained by selective absorption of BEC by the cancer cells which were present in abundance. Thus the bioavailablity of BEC to normal cells could be reduced.
  • ingestion of BEC into normal cells can occur by routes other than selective recognition by EEL's at high concentrations. For example, BEC at high concentrations may diffuse through the plasma membrane of the cell . Treatment of premalignant and malignant skin lesions with BEC in humans has also been studied.
  • Topical application of BEC has been observed to be effective for the treatment of lesions consisting of keratosis, basal cell carcinoma and squamous cell carcinoma.
  • Creams containing 10% and 0.005% BEC when applied topically showed complete clinical and histological regression when applied twice daily over treatment periods of up to about three months .
  • the duration of the treatment with the 0.005% BEC required for regression of the lesions was considerably longer than for the 10% BEC.
  • the treatment period required for the low concentration of BEC was about 13 to 14 weeks .
  • the extended duration of the treatment for the low concentration BEC formulations has a number of disadvantages.
  • the BEC formulation must be applied at regular intervals, typically twice a day, until clinical regression is observed.
  • Many patients find it difficult to comply with such a regime for up to 14 weeks. During this period, patients may experience an unacceptable amount of pain due to high salicyclic acid concentrations.
  • the lesions ulcerate and should be covered by a dressing. From a cosmetic and patient comfort perspective it would be desirable to be able to reduce the duration of treatment . Although such a reduction can be achieved by increasing the dose of BEC, this is undesirable in view of the toxicity of BEC.
  • Glycoalkaloids can undergo degradation in which the glycoside moiety or a saccharide unit thereof is cleaved from the alkaloid.
  • the glycoside moiety of the glycoalkaloid includes two or more saccharide units, there are a number of possible degradation products including free sugars such as monosaccharides, disaccharides and trisaccharides; the aglycone and mono and diglycosides .
  • free sugars refers to any sugar such as a mono, di, trisaccharide, oligosaccharide or polysaccharide or derivative thereof which is not bound to an alkaloid.
  • a medicinal composition comprising at least one compound which can interact with a target cell, the at least one compound being a glycoalkaloid of the general formula I:
  • either one of the dotted lines represents a double bond, and the other a single bond, or both represent single bonds ;
  • A represents a radical selected from the following radicals of general formulae (II) to (V) :
  • each of R 1 is a radical separately selected from the group consisting of hydrogen, amino, oxo and OR 4 ; each of R 2 is a radical separately selected from the group consisting of hydrogen, amino and OR 4 ' ' each of R 3 is a radical separately selected from the group consisting of hydrogen, alkyl and R 4 -alkylene; each of R 4 is a radical separately selected from the group consisting of hydrogen, carbohydrate and a carbohydrate derivative;" X" is a radical selected from the group comprising -CH 2 -, -0- and -NH 2 - ; wherein the compound includes at least one R 4 group in which R 4 is a carbohydrate or a derivative thereof ,- together with a pharmaceutically acceptable carrier, adjuvant, excipient and/or diluent, wherein the composition is essentially without free sugars of the type which inhibit the interaction between the at least one glycoalkaloid and a target cell .
  • Preferred carbohydrate radicals R 4 are glyceric aldehyde; glycerose; erythrose; threose; ribose; arabinose; xylose; lyxose; altrose; allose; gulose; mannose; glucose; idose; galactose; talose; rhamnose; dihydroxyactone; erythrulose; ribulose; xylulose; psicose; fructose; sorbose; tagatose; and other hexoses (C 6 H 12 0 6 ) ; heptoses (C 7 H 14 0 7 ) ; octoses (C 8 H 16 0 8 ) ; nanoses (C 9 H 18 0 9 ) ; decoses (C 10 H 20 O 10 ) ; deoxysugars with branched chains (eg.
  • apiose, hamamelose, streptose, cordycepose, mycarose and cladinose ; compounds wherein the aldehyde, ketone or hydroxyl groups have been substituted (eg. N- acetyl, acetyl, methyl, replacement of CH 2 0H) ; sugar alcohols; sugar acids; benzimidazoles; the enol salts of the carbohydrates; saccharinic acids; sugar phosphates.
  • the more preferred compounds are solasonine, solamargine, solanine and tomatine.
  • a preferred composition of the present invention is a solasodine glycoside composition which includes solasonine, solamargine and their di and monoglycosides in the same or similar proportion as the aforementioned BEC.
  • the composition of the present invention typically comprises naturally occurring glycoalkaloids extracted from a plant source.
  • the plant extract is treated to remove essentially all of any free sugars which can inhibit the efficacy of the glycoalkaloids prior to formulation of the composition of the present invention.
  • one or more free sugars do not inhibit the efficacy of the glycoalkaloids and do not need to be removed, typically all of the free sugars will be removed from the plant extract .
  • a method of preparing a glycoalkaloid preparation comprising at least one glycoalkaloid according to formula I, as hereinbefore defined, the method including extracting the at least one glycoalkaloid from a suitable plant material to form a crude extract, and removing essentially all free sugars from the crude extract .
  • the crude extract may be obtained by any suitable method.
  • the plant material is Solanum Sodomaeum a preferred method is to extract coarsely ground plant material with acetic acid.
  • the extract is filtered and the pH adjusted to about 9 to 10 to obtain a precipitate.
  • the precipitate may be dissolved in acetic acid and re-precipitated at high pH.
  • the precipitate is typically further extracted with ethanol to provide the solasodine glycoside mixture or BEC as a semicrystalline powder .
  • the free sugars may be removed from the plant extract by any suitable method.
  • a preferred method is to wash the crude extract in water or other suitable solvent.
  • the free sugars are removed to below detectable limits or are at least removed to a level below which an inhibitory effect can be detected.
  • the composition of the present invention is essentially without all free sugars. However, it will be appreciated that free sugars which do not inhibit the cytotoxicity of the glycoalkaloids may be present .
  • the composition of the present invention may also be formulated from a synthetic glycoalkaloid or a mixture of glycoalkaloids.
  • the synthetic glycoalkaloids would typically be treated prior to formulation of the composition to remove any sugars present as a result of glycoalkaloid degradation.
  • the glycoalkaloids in the composition of the present invention may also be obtained from chemical modification of naturally occurring glycoalkaloids.
  • the naturally occurring sugar moiety of the glycoalkaloid can be modified by removing or adding a saccharide unit or units. Suitable methods of carbohydrate modification are known and include chemical or enzymatic hydrolysis.
  • the sugar moiety may be completely removed and replaced with a different sugar moiety.
  • glycoalkaloids against target cells
  • EEL mediated endocytosis in which an EEL recognizes the sugar moiety of the glycoalkaloid and subsequent internalization of the EEL and glycoalkaloid.
  • a modified glycoalkaloid may be derived which is specific to that receptor. In this way a glycoalkaloid can be designed to target a desired cell type.
  • the products of glycoalkaloid degradation may also include the aglycone.
  • any aglycone is also removed prior to formulation of the therapeutic compositions of the present invention. Removal of the aglycone may be conducted by any suitable means and is typically removed by solvent extraction. Suitable solvents include the chlorinated hydrocarbon solvents and chloroform is particularly preferred.
  • the aforementioned sugar removal and if desired aglycone removal of stored glycoalkaloid be conducted immediately prior to formulation of the therapeutic compositions of the invention.
  • the composition is stabilized against glycoalkaloid degradation.
  • the composition is acidic and preferably includes acetic or lactic acid. The acidic conditions minimize degradation to produce free sugars .
  • sugar free glycoalkaloid preparations including the crystalline form may be prepared and then stored under stable conditions prior to formulation of the therapeutic composition of the present invention.
  • the sugar free preparation may be stored in an acidic solution and/or at low temperature.
  • a method of preparing a therapeutic composition which comprises a therapeutically effective amount of at least one glycoalkaloid according to formula I, as hereinbefore defined, the method including obtaining at least one glycoalkaloid, removing any free sugars from the glycoalkaloid and mixing the glycoalkaloid with a pharmaceutically acceptable stabilizer.
  • the amount of the glycoalkaloid present in the therapeutic composition of the present invention may depend on the dose rate, patient, the type of condition being treated and in the case of a tumor the type, size and position of the tumor to be treated.
  • a typical composition for the treatment of skin tumors would typically include between about 5 to about 0.001%, preferably about 0.005% solasodine glycosides.
  • the therapeutic composition of the present invention may be used in the treatment and control of conditions which may be treated or controlled by selective cellular destruction or modification. Such uses include the treatment or control of cancer, contraception, termination of pregnancy, removal of pathogenic organisms and removal of abnormal cellular growth. According to a further broad form of the present invention there is provided a method for the treatment or control of cancer, contraception, termination of pregnancy, removal of pathogenic organisms and removal of abnormal cellular growth in a mammal requiring such treatment, the method comprising administering to the mammal an effective amount of a medicinal composition or preparation of the present invention.
  • the medicinal composition of the present invention may be formulated in any suitable manner including injectable compositions, tablets, suppositories, capsules and topical formulations..
  • the formulation is a cream for topical administration or an injectable formulation.
  • the composition may be an injectable formulation for intraperitoneal or intralesional injection.
  • the injectable composition is administered in an amount of between about 50 to about 200 mg of sugar free glycoalkaloid composition per kg of tumour.
  • Animal and human studies show that successful treatment of some tumors and cancers may be accomplished with as few as two to four injections.
  • the injection may be given at one, two or three daily intervals, preferably the treatment is given twice, at day 1 and day 3. Treatment by injection may also be given in association with topical administration if desired or considered necessary.
  • the therapeutic composition of the present invention may also be used to diagnose skin conditions before such conditions can be detected by visual inspection.
  • Such diagnosis may be carried out by broadly applying a composition of the present invention to an area of skin to be tested.
  • the composition is left on the skin for a pre-determined period of time. During this time, any abnormal cells are selectively destroyed. This produces a detectable inflammation of the affected areas which may then be identified and treated.
  • a method of diagnosing a skin condition in a mammal, the skin condition being caused by the presence of abnormal cells wherein the method includes applying an effective amount of a composition or preparation of the present invention to an area of skin to be diagnosed, leaving the composition on the skin for a pre-determined period of time, removing the composition and detecting any change to any areas of skin.
  • the diagnostic method of the present invention is particularly suitable for diagnosing skin conditions of humans.
  • Typical conditions which may be diagnosed include Keratoses, basal cell carcinomas, squamous cell carcinomas, melanomas or other skin cancers.
  • a particularly preferred diagnostic composition is a solasodine glycoside mixture having about the same glycoside composition as BEC but without free sugars or the aglycone, solasodine. In trials conducted by the present inventor it has been observed that the normal healthy skin tissue is unaffected by the composition. This demonstrates the selectivity of glycoalkaloids for abnormal cells.
  • This method of diagnosis allows skin conditions to be detected and treated at an early stage, typically before the condition produces visible skin lesions.
  • the diagnosis can be conducted using very low concentrations of solasodine glycosides.
  • Figures 5 to 8 illustrate a sarcoid tumor in a horse, before ( Figure 5) , during ( Figures 6 and 7) and after ( Figure 8) treatment with a preferred composition of the present invention.
  • Figures 9 to 12 illustrate the stages in the treatment of a horse with a penile sarcoma before ( Figure 9) during ( Figures 10 and 11) and after ( Figure 12) treatment with a preferred composition of the present invention.
  • Figures 13 to 15 illustrate stages in the treatment of a human with squamous cell carcinoma before ( Figure 13) and during ( Figures 14 and 15) treatment.
  • a sugar free solasodine glycoside preparation was prepared according to the following:
  • the slurry is diluted with 3% acetic acid (pH
  • the solution is allowed to stand overnight without mixing.
  • the solution is subsequently filtered through a muslin cloth.
  • the filtrate is then subjected to a flow through centrifuge (3.5HP) at 1455 rpm.
  • the clear filtrate is heated to 50°C in a stainless steel double jacketed bowl.
  • Concentrated ammonia (L R Grade) is added until pH « 10. A precipitate is observed.
  • the precipitate is allowed to settle and cool (approx. 24 hrs) .
  • the supernatant is carefully decanted.
  • the precipitate is dissolved in 25L 3% aqueous acetic acid.
  • the solution is centrifuged through flow through centrifuge as above.
  • the supernatant is collected in an SS double jacketed bowl and heated to 50°C with continuous stirring (30 rpm, 30min) .
  • glycoalkaloids are re-precipitated by the addition of concentrated ammonia solution until pH « 10.
  • the solution is allowed to cool and the precipitate is allowed to settle (approx. 24 hrs) .
  • the supernatant is carefully decanted and the precipitate is washed with 50L water and allowed to settle for 24 hrs as before.
  • the supernatant is decanted and this procedure is repeated four times.
  • the precipitate is finally dissolved in 10L alcohol at 75°C and filtered whilst hot through Whatman No. 1 filter paper. The supernatant is dried at 50°C. This yields a fine, semicrystalline powder. The yield is 505g which is 1.01%.
  • any aglycone, solasodine is removed by washing the extract in chloroform.
  • the solasodine is soluble in the chloroform phase and the sugars are soluble in the aqueous phase.
  • the glycoalkaloids remain insoluble under all these conditions.
  • Cream formulations were prepared from the sugar free solasodine glycoside preparation from Example 1 as follows : Percentage Function Composition
  • Active ingredient Solasodine 0.005% w/w Antineoplast Glycosides (BEC)
  • BEC Antineoplast Glycosides
  • Salicylic acid 10.0% w/w Keratolytic Urea 5.0% w/w Keratolytic
  • Emulsifying wax, white soft paraffin, liquid paraffin, propylene glycol and water were used to provide a cream base of a suitable consistency and viscosity. Chlorocresol was included in the formulation as a preservative. Salicylic acid and urea were included as keratolytic agents and were considered to be excipients in the cream formulation because their primary function was to enhance the bioavailability of the active ingredient by clearing tissue from around the tumor, thus allowing a higher concentration of the active ingredients to reach the tumor. (The International Pharmaceutical Excipients Council definition of an excipient includes substances which are included in a drug delivery system to protect, support or enhance stability, bioavailability or patient acceptability and to enhance any other attribute of the overall safety and effectiveness of the drug during storage or use) .
  • Acetic or lactic acid was present in the final formulation because a 3% solution of acetic acid is used as the solvent for the active ingredients during the manufacturing process.
  • Purified water at 70°C, urea, chlorocresol and propylene glycol were added to a suitable stainless steel container and mixed for 2 minutes using a Silverson mixer ("Phase B") .
  • Phase A was slowed added to Phase B and thoroughly mixed using a Silverson mixer or follow- through homogenizer.
  • the mixture was allowed to cool to about 50°C and then the salicylic acid was added.
  • the freshly washed solasodine glycosides were dissolved in acetic acid or lactic acid solution and added to the cream, with mixing to ensure even dispersion.
  • the cream was allowed to cool to room temperature with occasional mixing to ensure an even, smooth texture.
  • the formulated cream had the following specifications :
  • solasodine glycoside preparation from Example 1 and creams from examples 2 and 3 were analyzed for hydrolysis products by MS and HPLC according to the following procedure.
  • Sample Preparation Standard was prepared in 50% CH 3 CN/H 2 0 at lmg/ml and lOOug/ml. Cream was prepared by dissolving lOOmg cream in 2ml methylene chloride, after centrifugation lOOul of the aqueous phase was removed and made up to 1ml with methanol . HPLC Conditions:
  • a Waters Alliance system was used consisting of a 2690 separations module, and 996 diode array detector.
  • a Micromass Waters Platform LCZ mass spectrometer was interfaced and the whole system was controlled by MassLynx chromatography software.
  • UV Detection was from 205nm to 320nm (205nm was extracted for alkaloids and 254 for salicylic acid and chlorocresol) Mass Spec full scans from 400-900 m/z were used for TIC chromatograms and single channels of 869 and 885 were used for quantitation.
  • Figures 1 to 4 illustrate HPLC spectra of unwashed ( Figures 1 and 3) and washed ( Figures 2 and 4) BEC respectively.
  • Figures 1 and 2 show that the unwashed BEC includes a number of further peaks. By comparison with Figures 3 and 4 , it can be seen that these peaks have been removed or significantly decreased upon washing with water and chloroform. These further peaks have been assigned to the various sugar degradation products of BEC.
  • Sodomaeum according to conventional extraction procedures .
  • the degradation of the BEC extract was estimated by measuring the change in solamargine and solasonine levels over time. Although degradation of BEC could also be measured by an increase in sugar levels, in practice HPLC analysis for solamargine and solasonine allowed a more quantative analysis to be conducted and was therefore chosen for this study. The results are illustrated in the following
  • mice with sarcoma 180 when treated with varying doses of 7mg and 8mg unwashed and washed BEC/kg. Washed BEC was prepared according to Example 1 and administered immediately after preparation.
  • Unwashed BEC was prepared in a similar manner but was not washed prior to use .
  • the unwashed crystalline BEC was stored under ambient conditions for about four weeks prior to administration to the mice.
  • the % degradation over the four weeks can be estimated to be between less than 4 to 5% for solamargine and solasonine respectively.
  • BEC should be washed prior to formulation even after storage for even short periods of time (such as about four days) .
  • mice with sarcoma 180 were treated with 7mg/kg doses given on consecutive days. The results are shown in the following table. COMPOUND DOSE NUMBER SURVIVAL ANIMALS
  • Each value is the mean ⁇ S D obtained in groups of twelve tumour - bearing mice treated intraperitoneally 0.5h after tumour implantation (5 x 10 5 cells/mouse).
  • a The criterion of survival was taken as 56 days because it was shown that if the treatment was effective against sarcoma 180 for this period, the mice then had a normal life span (approximately 3 years) .
  • % survival for animals treated with the washed BEC was superior when compared with animals treated with an equivalent dose of unwashed BEC.
  • % survival rate for four doses of unwashed BEC is 50% as compared with 100% for washed BEC.
  • the effect of washed and unwashed BEC or human ovarian cancer cells was compared.
  • the washed and unwashed BEC were prepared as described for Example 8.
  • BEC solasodine glycosides
  • Example 9 A patient with no visible lesions on the face had a cream as prepared in Example 2 applied to the skin of the face. The cream was left for 30 minutes before being washed away. The patient's face was then examined. Areas of redness were noted which were identified as pre-malignant or malignant skin lesions in the very early stages of development . The affected areas of skin were subsequently treated with the same cream.
  • Example 11 A human patient was diagnosed with an intra epithelial penile tumour. The prognosis was that no treatment was available and that amputation was the only option. The patient commenced treatment with the cream as prepared in Example 2 and was applied to the tumour twice daily. Necrosis of the tumour was observed to occur shortly after treatment commenced. Within six weeks, the patient was observed to be free of the tumour. Example 11
  • Formulation A sugar and aglycone free solasodine glycoside preparation which was prepared according to Example 1 in DMSO. DMSO is used for its aprotic characteristics and because when pure is sterile.
  • Figures 5, 6, 7 and 8 show an example of a sarcoid tumor in a horse, before, during and after treatment.
  • Figure 5 illustrates the sarcoma before treatment .
  • Figures 6 and 7 show the sarcoma after injection with the above composition. Necrosis of the sarcoma can be seen.
  • Figure 4 shows that the sarcoma has fully regressed after treatment.
  • Figures 9 to 12 show a further example of the treatment of a horse with a penile tumor before, during and after treatment.
  • Figure 9 shows the horse prior to treatment.
  • the horse was anesthetized and the tumor injected with the above formulation.
  • Figure 10 shows the response of the tumor to the composition. The tumor then separated entirely and fell off as shown in Figure 11.
  • Figure 12 illustrates the penis after the treatment.
  • Figures 13, 14 and 15 show an example of the treatment of a human SCC.
  • Figure 13 shows the SCC located on the patient's scalp. The patient was treated with a single injection and recovery of the SCC shortly after treatment occurred as illustrated in Figures 14 and 15.
  • composition of the present invention was successful in the treatment of solid sarcoids in animals and SCC in humans. During treatment, necrosis of the lesion was observed to begin almost immediately after injection.
  • the dosages of the compositions in the above examples is lOOmg of solasodine glycosides per 1kg tumor.
  • a typical tumor is lOOg such that a typical injection contains lOmg solasodine glycosides. This dose for a 500kg horse corresponds to 0.02mg/kg body weight.
  • a therapeutic composition of the present invention provides a suprising and unexpected improvement in efficacy of glycoalkaloids in the treatment of cancers and tumors.
  • This increase in efficacy allows disease conditions to be treated with dosages which are well below the threshold level of toxicity for normal cells.
  • This is advantageous for the patient and also allows the inventive compositions to be used as diagnostic tools.
  • the improved efficacy enables the duration of treatments to be reduced and total dosages to be decreased. This is advantageous for the overall safety and comfort of the patient and also provides a superior treatment regime in terms of cost effectiveness.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Epidemiology (AREA)
  • Molecular Biology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Dermatology (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

Cette invention se rapporte à une composition médicinale comprenant au moins un composé pouvant interagir avec une cellule cible, ce ou ces composés étant constitués par un glycoalcaloïde, représenté par la formule générale (I), où cette composition est essentiellement exempte de sucres libres empêchant l'interaction entre le ou les glycoalcaloïdes et une cellule cible.
PCT/AU2000/000300 1999-04-09 2000-04-10 Compositions medicinales et leur procede de preparation WO2000061153A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
AU35454/00A AU779512B2 (en) 1999-04-09 2000-04-10 Medicinal compositions and their method of preparation
EP00913972A EP1181022A4 (fr) 1999-04-09 2000-04-10 Compositions medicinales et leur procede de preparation
CA002369272A CA2369272A1 (fr) 1999-04-09 2000-04-10 Compositions medicinales et leur procede de preparation

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
AUPP9686 1999-04-09
AUPP9686A AUPP968699A0 (en) 1999-04-09 1999-04-09 Therapeutic compositions and method for their preparation

Related Child Applications (2)

Application Number Title Priority Date Filing Date
US09958333 A-371-Of-International 2001-12-20
US10/752,095 Continuation US20040220115A1 (en) 1999-04-09 2004-01-07 Medicinal compositions and their method of preparation

Publications (1)

Publication Number Publication Date
WO2000061153A1 true WO2000061153A1 (fr) 2000-10-19

Family

ID=3813885

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/AU2000/000300 WO2000061153A1 (fr) 1999-04-09 2000-04-10 Compositions medicinales et leur procede de preparation

Country Status (4)

Country Link
EP (1) EP1181022A4 (fr)
AU (1) AUPP968699A0 (fr)
CA (1) CA2369272A1 (fr)
WO (1) WO2000061153A1 (fr)

Cited By (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003029269A1 (fr) * 2001-09-28 2003-04-10 Glycomed Sciences Limited Procede d'extraction de solvant
WO2003030884A2 (fr) * 2001-10-09 2003-04-17 Glycomed Sciences Limited Utilisation de solasonine
WO2003030915A1 (fr) * 2001-10-09 2003-04-17 Glycomed Sciences Limited Utilisation de solamargine pour le traitement de tumeurs de la peau
WO2004002998A1 (fr) * 2002-06-28 2004-01-08 Solbec Pharmaceuticals Limited Procede de separation de triglycoalcaloïdes
WO2004062675A1 (fr) * 2003-01-15 2004-07-29 Solbec Pharmaceuticals Limited Procede de modulation de il-6
EP1508334A1 (fr) * 2003-08-22 2005-02-23 G & E Herbal Biotechnology Co., Ltd. Extrait soluble dans l'eau de solanum, procédé de préparation et composition pharmaceutique de l'extrait hydrosoluble
US6984725B2 (en) 2002-06-28 2006-01-10 Solbec Pharmaceuticals Ltd. Method for the separation of triglycoalkaloids
US7078063B2 (en) 2003-06-05 2006-07-18 G & E Herbal Biotechnology Co., Ltd. Water soluble extract from plant of Solanum genus and the preparation process thereof, and pharmaceutical composition containing the water soluble extract
WO2006092017A1 (fr) * 2005-03-02 2006-09-08 Solbec Pharmaceuticals Limited Combinaisons d'un glycoalcaloïde et d'un agoniste de tlr et diverses applications desdites combinaisons
US7138427B2 (en) 1999-03-26 2006-11-21 Phytopharm Plc. 5-β-sapogenin and pseudosapogenin derivatives and their use in the treatment of dementia
US7906493B2 (en) 2003-12-22 2011-03-15 Btg International Limited Core 2 GlcNAc-T inhibitors
US7964223B2 (en) * 2005-09-27 2011-06-21 University Of Kentucky Research Foundation Berry preparations and extracts
US7998943B2 (en) 2005-07-06 2011-08-16 Btg International Limited Core 2 GlcNAc-T inhibitors III
US8197794B2 (en) 2003-12-22 2012-06-12 Ms Therapeutics Limited Core 2 GlcNAc-T inhibitors
EP2520303A1 (fr) * 2011-05-04 2012-11-07 G & E Herbal Biotechnology Co., Ltd. Composition pharmaceutique anti-verrues
EP2522357A1 (fr) * 2011-05-12 2012-11-14 G & E Herbal Biotechnology Co., Ltd. Traitement et/ou prévention de l'inflammation et des lésions cutanées causées par la lumière et photoprotection de la peau avec un extrait soluble dans l'eau d'une plante du genre solanum
US8609633B2 (en) 2005-07-06 2013-12-17 Ms Therapeutics Limited Core 2 GlcNAc-T inhibitors
WO2017147659A1 (fr) 2016-03-03 2017-09-08 Bill Elliot Cham Combinaisons de glycoalcaloïdes et leurs diverses utilisations
RU2795113C1 (ru) * 2016-03-03 2023-04-28 Билл Эллиот ЧЭМ Комбинации гликоалкалоидов и различные их применения

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3520793B1 (fr) * 2011-06-06 2023-03-29 University of Iowa Research Foundation Substances et compositions à utiliser dans des procédés d'inhibition de l'amyotrophie
CN105192820A (zh) * 2015-08-20 2015-12-30 泰山医学院 一种降血脂蛹虫草多糖饮料及其制备方法

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU540812B2 (en) * 1979-05-02 1984-12-06 Aruba Qld Pty. Ltd. Steroid alkaloids
AU654474B2 (en) * 1990-01-18 1994-11-10 Cura Nominees Pty Ltd Glycoalkaloids

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU540812B2 (en) * 1979-05-02 1984-12-06 Aruba Qld Pty. Ltd. Steroid alkaloids
AU654474B2 (en) * 1990-01-18 1994-11-10 Cura Nominees Pty Ltd Glycoalkaloids

Non-Patent Citations (12)

* Cited by examiner, † Cited by third party
Title
CAHM B.E. ET AL., CANCER LETTERS,, vol. 59, 1991, pages 183 - 192, XP002950614 *
CHAM B.E. ET AL., CANCER LETTERS,, vol. 55, 1990, pages 221 - 225, XP002950613 *
CHAM B.E., ASIA PACIFIC JOURNAL OF PHARMACOLOGY,, vol. 9, 1994, pages 113 - 118, XP002950616 *
CHAM B.E., DRUGS OF THE FUTURE,, vol. 13, no. 8, 1988, pages 714 - 716, XP002950611 *
CHAM BILL E. ET AL., CANCER LETTERS,, vol. 36, 1987, pages 111 - 118, XP002950615 *
CHAM BILL E. ET AL.: "Antitumor ..... Solanum Sodomaeum", PLANTA MEDICA,, vol. 53, no. 1, 1987, pages 34 - 36, XP002950610 *
CHATAING B. ET AL., PLANTA MEDICA,, vol. 64, 1998, pages 31 - 36, XP002950618 *
DAUNTER B. ET AL., CANCER LETTERS,, vol. 55, 1990, pages 209 - 220, XP002950612 *
ELICH F, CRIBB A B, CRIBB J W: "WILD MEDICINE IN AUSTRALIA, PASSAGE", WILD MEDICINE IN AUSTRALIA, XX, XX, 1 January 1988 (1988-01-01), XX, pages 198/199, XP002950609 *
KUPCHAN S.M., PURE AND APPLIED CHEMISTRY,, vol. 21, 1965, pages 227 - 246, XP002950608 *
LI-CHING CHANG ET AL., BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,, vol. 242, 1998, pages 21 - 25, XP002950617 *
See also references of EP1181022A4 *

Cited By (31)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7507720B2 (en) 1998-03-26 2009-03-24 Phytopharm Plc 5-Beta-sapogenin and pseudosapogenin derivatives and their use in the treatment of dementia
US7138427B2 (en) 1999-03-26 2006-11-21 Phytopharm Plc. 5-β-sapogenin and pseudosapogenin derivatives and their use in the treatment of dementia
WO2003029269A1 (fr) * 2001-09-28 2003-04-10 Glycomed Sciences Limited Procede d'extraction de solvant
US7479290B2 (en) 2001-09-28 2009-01-20 Glycomed Sciences Limited Solvent extraction process
WO2003030884A3 (fr) * 2001-10-09 2003-09-04 Glycomed Sciences Ltd Utilisation de solasonine
WO2003030915A1 (fr) * 2001-10-09 2003-04-17 Glycomed Sciences Limited Utilisation de solamargine pour le traitement de tumeurs de la peau
WO2003030884A2 (fr) * 2001-10-09 2003-04-17 Glycomed Sciences Limited Utilisation de solasonine
US6984725B2 (en) 2002-06-28 2006-01-10 Solbec Pharmaceuticals Ltd. Method for the separation of triglycoalkaloids
WO2004002998A1 (fr) * 2002-06-28 2004-01-08 Solbec Pharmaceuticals Limited Procede de separation de triglycoalcaloïdes
WO2004062675A1 (fr) * 2003-01-15 2004-07-29 Solbec Pharmaceuticals Limited Procede de modulation de il-6
US7078063B2 (en) 2003-06-05 2006-07-18 G & E Herbal Biotechnology Co., Ltd. Water soluble extract from plant of Solanum genus and the preparation process thereof, and pharmaceutical composition containing the water soluble extract
JP2007145849A (ja) * 2003-06-05 2007-06-14 G & E Herbal Biotechnology Co Ltd Ｓｏｌａｎｕｍ属の植物由来の水溶性抽出物およびそれらの調製方法、ならびに水溶性抽出物を含む薬学的組成物
EP1508334A1 (fr) * 2003-08-22 2005-02-23 G & E Herbal Biotechnology Co., Ltd. Extrait soluble dans l'eau de solanum, procédé de préparation et composition pharmaceutique de l'extrait hydrosoluble
US7906493B2 (en) 2003-12-22 2011-03-15 Btg International Limited Core 2 GlcNAc-T inhibitors
US8197794B2 (en) 2003-12-22 2012-06-12 Ms Therapeutics Limited Core 2 GlcNAc-T inhibitors
WO2006092017A1 (fr) * 2005-03-02 2006-09-08 Solbec Pharmaceuticals Limited Combinaisons d'un glycoalcaloïde et d'un agoniste de tlr et diverses applications desdites combinaisons
US8609633B2 (en) 2005-07-06 2013-12-17 Ms Therapeutics Limited Core 2 GlcNAc-T inhibitors
US7998943B2 (en) 2005-07-06 2011-08-16 Btg International Limited Core 2 GlcNAc-T inhibitors III
US7964223B2 (en) * 2005-09-27 2011-06-21 University Of Kentucky Research Foundation Berry preparations and extracts
EP2520303A1 (fr) * 2011-05-04 2012-11-07 G & E Herbal Biotechnology Co., Ltd. Composition pharmaceutique anti-verrues
US8614196B2 (en) 2011-05-12 2013-12-24 G & E Herbal Biotechnology Co., Ltd. Treatment and/or prevention of inflammation and cutaneous photodamage and photoprotection of the skin with a water-soluble extract from plant of Solanum genus
JP2012236815A (ja) * 2011-05-12 2012-12-06 G & E Herbal Biotechnology Co Ltd ナス属（ｓｏｌａｎｕｍｇｅｎｕｓ）植物からの水溶性抽出物での炎症及び皮膚の光損傷の治療及び／又は予防、並びに皮膚の光防護
EP2522357A1 (fr) * 2011-05-12 2012-11-14 G & E Herbal Biotechnology Co., Ltd. Traitement et/ou prévention de l'inflammation et des lésions cutanées causées par la lumière et photoprotection de la peau avec un extrait soluble dans l'eau d'une plante du genre solanum
AU2012200390B2 (en) * 2011-05-12 2014-02-20 G & E Herbal Biotechnology Co., Ltd. Treatment and/or prevention of inflammation and cutaneous photodamage and photoprotection of the skin with a water-soluble extract from plant of solanum genus
KR101397107B1 (ko) 2011-05-12 2014-05-19 지앤이 허벌 바이오테크놀로지 씨오., 엘티디. 솔라눔 속 식물의 수용성 추출물에 의한 염증 및 피부의 광손상 치료 및/또는 예방 및 피부의 광보호
WO2017147659A1 (fr) 2016-03-03 2017-09-08 Bill Elliot Cham Combinaisons de glycoalcaloïdes et leurs diverses utilisations
KR20180117681A (ko) * 2016-03-03 2018-10-29 빌 엘리엇 참 글리코알칼로이드 조합 및 그의 다양한 용도
JP2019510813A (ja) * 2016-03-03 2019-04-18 エリオット チャム，ビル グリコアルカロイド組み合わせ及びそれらの多様な使用
KR102485909B1 (ko) * 2016-03-03 2023-01-06 빌 엘리엇 참 글리코알칼로이드 조합 및 그의 다양한 용도
JP7202188B2 (ja) 2016-03-03 2023-01-11 エリオット チャム，ビル グリコアルカロイド組み合わせ及びそれらの多様な使用
RU2795113C1 (ru) * 2016-03-03 2023-04-28 Билл Эллиот ЧЭМ Комбинации гликоалкалоидов и различные их применения

Also Published As

Publication number Publication date
EP1181022A4 (fr) 2004-02-11
EP1181022A1 (fr) 2002-02-27
AUPP968699A0 (en) 1999-05-06
CA2369272A1 (fr) 2000-10-19

Similar Documents

Publication Publication Date Title
EP1181022A1 (fr) Compositions medicinales et leur procede de preparation
EP2346518B1 (fr) Procede d' extraction de glucosides cardiotoniques à partir de nerium oleander et compositions
JP3530525B2 (ja) 表皮の再生を促進し、毛髪の再生を刺激し、または毛髪の喪失を遅らせる化粧組成物または医薬組成物、とりわけ皮膚科組成物の調製のためのメジカゴサポニンの使用
US20040220115A1 (en) Medicinal compositions and their method of preparation
EP0727217A2 (fr) Compositions pharmaceutiques et cosmétiques contenant ellagitannin du type god comme principe actif
WO1996000561A1 (fr) Produit stimulant la pousse des cheveux
US5556866A (en) Hair restorer and its preparation
KR19990014205A (ko) 플라보노이드 및 파피아 가공물을 함유한 조성물
US7935672B2 (en) Derivatives of genkwanin and sakuranetin, cosmetic and therapeutic use thereof and preparation method of same
HU228799B1 (en) Tanacetum parthenium extract
CN108883123B (zh) 配糖生物碱组合及其各种用途
US6939859B1 (en) Skin preparations for external use
CA2312856C (fr) Utilisation d'extraits de ginkgo biloba pour preparer un medicament
AU779512B2 (en) Medicinal compositions and their method of preparation
US11427611B2 (en) Use of steroidal glycosides, pharmaceutical formulations, use of Furcraea foetida plant extracts, process for producing Furcraea foetida plant extracts and method for treating skin disorders
RU2795113C1 (ru) Комбинации гликоалкалоидов и различные их применения
KR102014685B1 (ko) 골담초 유래 화합물 및 이를 함유하는 피부 미백용 조성물
CN110840878B (zh) 一种用于治疗银屑病的化合物及其制备方法
JP3182047B2 (ja) 黒砂糖より抽出された成分を含む化粧料組成物
JP3246683B2 (ja) 細胞増殖促進物質及びそれを含有する組成物
JP3182044B2 (ja) 黒砂糖より抽出された成分を含む化粧料組成物
WO2003022231A1 (fr) Compositions pour stimuler et favoriser la pousse des cheveux
JP2001302487A (ja) 美白化粧料

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY CA CH CN CR CU CZ DE DK DM DZ EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
ENP Entry into the national phase

Ref document number: 2369272

Country of ref document: CA

Kind code of ref document: A

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: 2000913972

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 35454/00

Country of ref document: AU

WWE Wipo information: entry into national phase

Ref document number: 09958333

Country of ref document: US

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

WWP Wipo information: published in national office

Ref document number: 2000913972

Country of ref document: EP

NENP Non-entry into the national phase

Ref country code: JP

WWG Wipo information: grant in national office

Ref document number: 35454/00

Country of ref document: AU