Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
• • 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
  • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
  • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
  • A61K31/404—Indoles, e.g. pindolol
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
  • A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/10—Dispersions; Emulsions
  • A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/20—Pills, tablets, discs, rods
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/20—Pills, tablets, discs, rods
  • A61K9/2004—Excipients; Inactive ingredients
  • A61K9/2022—Organic macromolecular compounds
  • A61K9/2031—Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyethylene oxide, poloxamers
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P13/00—Drugs for disorders of the urinary system
  • A61P13/08—Drugs for disorders of the urinary system of the prostate
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders

Definitions

• the invention relates to the field of pharmacy and relates to new pharmaceutical compositions for oral delivery of 3,3'-diindolylmethane (DIM) and methods for treating diseases using them.
• DIM 3,3'-diindolylmethane
• 3,3'-diindolylmethane (DIM) has a wide range of biological activities, which allows us to consider it as a very promising pharmacologically active compound.
• 3,3'-diindolylmethane (DIM) is the main oligomeric product of indole-3-carbinol (13 C), which shows a pronounced selective activity against transformed cells of different origin (Aggarwal BB, Ichikawa N. (2005) Molecular targets and anticancer potential of indole-3 -carbinol and its derivatives. Cell Cycle, 4 (9), 1201-1215).
• the hormone-sensitive cells of the prostate gland have shown the ability of DIM to competitively bind to androgen receptors, thus inhibiting their translocation into the nucleus, followed by activation of gene transcription, as well as the expression of the promoter of the gene encoding the prostate-specific antigen PSA.
• Protein PSA protease
• DIM is extremely similar to the known Casodex synthetic antiandrogen (Le NT, Schaldach CM, Bjeldanes LF.
• Plant-derived 3,3'-diindolylmethane is a strong androgen antagonist in human prostate cancer cells, J. Biol. Chem., 278, 21136-21145), which, however, unlike DIM, promotes translocation of androgen receptors into the nucleus (Masiello D, Cheng S, Bubley GJ et al. (2002) Bicalutamide functions as an androgen receptor antagonist by assembly of a transcriptionally inactive receptor, J. Biol Chem., 277, 26321-26326).
• NF- ⁇ nuclear transcription factor NF- ⁇
• proliferative antiapoptotic
• angiogenic migratory
• invasive cellular activities performing the final stage of signaling cascades induced by growth factors and cytokines.
• the key point is the translocation of the active factor into the nucleus and the activation of transcription of the genes responsible for these processes. It has been found that in vitro DIM (Rahman KM, Ali S, Aboukameel A et al.
• DIM DIM-induced apoptosis of cervical HPV-infected human keratinocytes in vitro.
• DIM showed several times greater efficiency than I3C.
• the LD 50 value was 50-60 ⁇ M for DIM and 200 ⁇ M for I3C, respectively, but like its metabolic precursor (I3C), DIM did not cause apoptotic changes in normal (untransformed) keratinocytes (Chen DZ, Qi M, Auborn K, Carter TH.
• DIM DIM- its immunomodulatory activity.
• DIM has been shown to stimulate IFNY-dependent signaling cascades by activating expression of IFNy receptors as well as other IFN-responsive regulatory proteins.
• Oral dosing of DIM-based drugs is most preferred since it contains a number of advantages over other dosing methods, such as the greatest comfort for the patient, the possibility of flexible treatment regimens, as well as the effectiveness of the cost of treatment.
• the bioavailability of DIM by oral dosing is severely limited due to its very low solubility and low absorption efficiency in the small intestine.
• DIM typically exhibits low solubility in body fluids and has very limited ability to penetrate barrier membranes.
• the ability of this compound to bind to plasma proteins and to participate in various non-specific interactions in the bloodstream is well known, which significantly reduces the efficiency of its delivery to the focus of the disease.
• TPGS pegylated vitamin E
• an object of the present invention is to solve the problem of improved DIM delivery.
• the pharmaceutical composition for oral administration contains 3,3'-diindolylmethane as an active ingredient and a target additive, and it contains as a target additive a block copolymer of oxyethylene and oxypropylene, in which the content of the hydrophobic block is less than 50 wt.%, And the molecular weight is hydrophilic block is 2250 Yes or more, with a ratio of the active component and the selected block copolymer of 10: 1-2: 1.
• the pharmaceutical composition preferably contains as a block copolymer of oxyethylene and oxypropylene Pluronic F127
• the pharmaceutical composition may further comprise Pluronic L10.
• the pharmaceutical composition may also further comprise a pharmaceutically acceptable carrier.
• the pharmaceutical composition may be a tablet, lyophilized powder, suspension or capsule. LIST OF DRAWINGS
• composition 1 Composition 1 - DIM (control)
• composition 3 Pluronic F127, Pluronic L10 and DIM
• composition 1 Composition 1 - DIM (control),
• composition 3 freeze-dried solution of Pluronic F127, Pluronic L10 and DIM
• FIG. 3 data from a morphological study in patients with prostate adenoma (AP) and intraepithelial neoplasia of the prostate (IDU) before and after treatment:
• FIG. 4 data of immunohistochemical analysis, investigated such indicators as growth factors IGF and EGF, regulatory factor TGF-D, before and after prescribing:
• Group I - (4 patients) took the claimed pharmaceutical composition with DIM
• Group II - (4 patients) took a pharmaceutical composition with crystalline DIM.
• the block copolymers of oxyethylene and oxypropylene are also known as Pluronic or Poloxamer.
• the hydrophobic-hydrophilic properties of Pluronic and their ability to solubilize water-insoluble compounds are determined by the size and composition of the polyoxyethylene (hydrophilic) and polyoxypropylene (hydrophobic) blocks.
• Table 1 summarizes the structural properties of the various Pluronics.
• block copolymers are widely used in pharmaceutical and cosmetic compositions, including to increase the solubility of hydrophobic water-insoluble compounds (Foster B, Cosgrove T, Hammouda B (2009) Pluronic triblock copolymer systems and their interactions with ibuprofen, Langmuir, 25 (12), 6760-6766) requires an individual decision on their use for each specific medicine.
• block copolymers of oxyethylene and oxypropylene to enhance the oral bioavailability of the compounds, it is associated with modulation of the activity of P-glycoprotein and, accordingly, they are proposed in compositions where the active substances are substrates of P-glycoprotein (Kabanov et al. US patent 6.277.410 Copolymer compositions for oral delivery) and some forms of MRP (Miller DW, Batrakova EV, Kabanov AV (1999) Inhibition of multidrug resistance-associated protein (MRP) functional activity with pluronic block copolymers, Pharm Res, 16 (3), .396- 401). Similar properties have also been demonstrated for other polymeric surface active compounds.
• Example 8 A direct experiment described below (Example 8) showed that DIM does not increase the uptake of rhodamine by 123 cells expressing P-glycoprotein, which indicates that DIM is not a substrate of this transporter. This is in good agreement with the results of the above attempt to use the highly active TPGS P-glycoprotein modulator to increase the bioavailability of DIM, which did not lead to a significant improvement in this parameter. However, the use of a number of surface-active polymers described by this election has led to a significant increase in the oral bioavailability of the drug.
• the bioavailability of DIM also depends on the molecular weight of the hydrophilic block. Higher bioavailability is provided by block copolymers of oxyethylene and oxypropylene, in which the content of the hydrophobic block is less than 50 wt.%, And the molecular weight of the hydrophilic block is 2250 Da or more.
• the most effective was Pluronic F127, allowing to obtain stable aqueous dispersions containing more than 3 mg / ml DIM. Oral administration of such a DIM composition to rats allowed a significant (more than 5-fold) increase in its bioavailability.
• the ratio of the active component and the selected block copolymer may vary depending on the desired duration of release and is on average 10: 1-2: 1.
• the most optimal ratio of Pluronic F127 and DIM was 6: 1.
• Pluronic L10 hydrophilic block content of about 40% and total molecular weight of about 3200
• Pluronic F 127 hydrophilic block content of about 40% and total molecular weight of about 3200
• Pluronic P85 and Pluronic L61 did not have a similar effect.
• compositions according to the invention can be obtained, for example, by co-dissolving or separately dissolving the components in suitable solvents, such as water, alcoholic or aqueous-alcoholic solutions, then mixing the resulting solutions in the required proportions.
• suitable solvents such as water, alcoholic or aqueous-alcoholic solutions
• the resulting solutions may optionally be dried to obtain a solid dosage form. Drying is carried out by any technologically suitable method or a combination thereof, including but not limited to methods such as rotary evaporator or speed evaporation, freeze drying, stream drying.
• Finished dosage forms can be obtained by tabletting the dried compositions using the necessary excipients, for example, sodium stearate, lactose, cellulose derivatives, etc.
• Finished dosage forms can be obtained by packaging the dried compositions in capsules, for example, hard-gelatin capsules.
• compositions containing an effective amount of DIM can be used to treat various diseases.
• the described compositions are proposed for the treatment of proliferative diseases such as uterine fibroids, adenomyosis, and hyperplastic diseases of the prostate gland.
• proliferative diseases such as uterine fibroids, adenomyosis, and hyperplastic diseases of the prostate gland.
• the effective amount of diindolylmethane needed for treatment and prophylaxis may vary depending on the type and severity of the disease, the age and condition of the patient, and can be determined in each case by the attending physician. In this case, the doses used are in the range from 2 to 2000 mg per day.
• Rhodamine 123 Rhodamine 123 (R123) by MESSA / DX cells expressing membrane P-glycoprotein (P-gp) was studied.
• P-gp negative MESSA / DX cells were used as a control. Cells were placed in a 96-well a plateau at a concentration of 40,000 cells per well. After 24 hours of incubation, R123 was added to the cells at a concentration of 3 ⁇ m. known P-gp inhibitor After incubation, the solution was removed and the cells were washed 3 times with chilled phosphate buffer. After that, Rhodamine fluorescence was measured in cell samples. All experiments were performed three times.
• MESSA / DX cells expressing membrane P-glycoprotein (P-gp) weakly absorb rhodamine compared to P-gp negative MESSA cells.
• Verapamil a known inhibitor of P-gp in a dose-dependent manner, increases the accumulation of R123 in MESSA / DX cells, but does not affect its accumulation in MESSA cells.
• DIM at a concentration of 500 ⁇ m does not affect the accumulation of R123 in any of the cell lines, which clearly indicates that DIM is not a substrate of P-gp. ''
• compositions were given to animals orally in the form of aqueous dispersions with a final concentration of DIM of 3 mg per ml.
• Crystalline DIM was given as a suspension of 15 mg DIM in 5 ml of 0.5% methylcellulose in distilled water.
• DIM formulations were administered to animals at a dosage of 60 mg per kg of body weight. After receiving the drugs at various time intervals (15, 30, 45 minutes and 1, 2, 4, 6 and 24 hours), blood samples were collected from animals. Right after blood samples were centrifuged, and the separated plasma was frozen and stored at -80 ° C.
• Isoflurane (Bimeta-MTC, Animal Health Inc. Cambridge, ON, Canada). Blood was drawn from the jugular vein into heparin tubes, which were immediately placed on ice for 5 to 10 minutes. Then, the blood was centrifuged to separate it from the plasma. Samples with plasma were frozen and stored at -80 ° C.
• Samples with plasma were thawed, centrifuged, aliquots of 100 ⁇ l were packaged in plastic tubes. Next, the samples were extracted twice with 750 ⁇ l of methyl tert-butyl ether for 2 min stirring at 180 ° C. Samples were centrifuged at 10,000 for 10 minutes. Supernatants were separated and transferred to glass tubes. The organic phase was evaporated with nitrogen at 50 ° C until completely dried. The dried samples were stored at a temperature of -80 ° C. The test samples were diluted in 15 ⁇ l of acetonitrile and 85 ⁇ l of the mobile phase. Then aliquots in a volume of 20 ⁇ l were analyzed using the HPLC method.
• Ci8 reverse phase columns 50 x 4.6 mm, Symmetry / shield 3.5 ⁇ m (sorbent, grain size in ⁇ m), 30 ° C, flow rate 1.5 ml / min, injection volume 20 ⁇ l, at 280 nm.
• Example 10 The study of the clinical efficacy of the pharmaceutical composition DIM obtained according to Example 2.
• composition with DIM according to the invention (group I) - 2 capsules 12 times a day.
• composition with crystalline DIM (group II) - 2 capsules / 2 times a day To assess the clinical efficacy, morphological effects and safety of the drugs, 34 patients with prostate adenoma (AP) and prostate intraepithelial neoplasia (IDU) were examined and treated. Group I (18 patients) took the proposed pharmaceutical composition with DIM 2 capsules 2 times a day, group II (16 patients) took the pharmaceutical composition with crystalline DIM 2 capsules 2 times a day.
• Residual urine no more than 200 ml
• the prostate volume is more than 25 cubic meters. cm;
• Immunohistochemical analysis was carried out in two groups of patients, consisting of four people from the group taking the pharmaceutical composition with DIM in the new form (group I) and the pharmaceutical composition with crystalline DIM (group II), respectively.
• group I the pharmaceutical composition with DIM
• group II the pharmaceutical composition with crystalline DIM
• IGF and EGF growth factors
• TGF-D regulatory factor-D
• the obtained data of a significant decrease in IGF and EGF growth factors, an increase in the level of TGF-D in the group receiving the claimed pharmaceutical composition with DIM indicate the effect of the active ingredients of the drug on the main signaling mechanisms of pathological cell proliferation, as well as on the induction of apoptosis of transformed cells.
• the pharmaceutical composition with DIM in the new formulation has antiproliferative activity in patients with prostate adenoma and prostatic intraepithelial neoplasia.
• the pharmaceutical composition with DIM in the new formulation is a safe tool in the treatment of APA and IDUs in view of the absence of side effects and adverse events during treatment.

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• 2009
  • 2009-09-18 RU RU2009134872/15A patent/RU2409363C9/ru active
• 2010
  • 2010-09-07 WO PCT/RU2010/000487 patent/WO2011034465A1/ru not_active Ceased
  • 2010-09-07 US US13/377,261 patent/US8791150B2/en not_active Expired - Fee Related
  • 2010-09-07 CN CN2010800524064A patent/CN102711755A/zh active Pending
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