CN117447476A - Temozolomide-caffeic acid medicine co-crystal and preparation method thereof - Google Patents
Temozolomide-caffeic acid medicine co-crystal and preparation method thereof Download PDFInfo
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- CN117447476A CN117447476A CN202311409216.9A CN202311409216A CN117447476A CN 117447476 A CN117447476 A CN 117447476A CN 202311409216 A CN202311409216 A CN 202311409216A CN 117447476 A CN117447476 A CN 117447476A
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- 229940074360 caffeic acid Drugs 0.000 title claims abstract description 141
- 239000013078 crystal Substances 0.000 title claims abstract description 126
- 239000003814 drug Substances 0.000 title claims abstract description 61
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- QAIPRVGONGVQAS-DUXPYHPUSA-N trans-caffeic acid Chemical compound OC(=O)\C=C\C1=CC=C(O)C(O)=C1 QAIPRVGONGVQAS-DUXPYHPUSA-N 0.000 claims abstract description 102
- BPEGJWRSRHCHSN-UHFFFAOYSA-N Temozolomide Chemical compound O=C1N(C)N=NC2=C(C(N)=O)N=CN21 BPEGJWRSRHCHSN-UHFFFAOYSA-N 0.000 claims abstract description 57
- 229960004964 temozolomide Drugs 0.000 claims abstract description 57
- ACEAELOMUCBPJP-UHFFFAOYSA-N (E)-3,4,5-trihydroxycinnamic acid Natural products OC(=O)C=CC1=CC(O)=C(O)C(O)=C1 ACEAELOMUCBPJP-UHFFFAOYSA-N 0.000 claims abstract description 51
- 235000004883 caffeic acid Nutrition 0.000 claims abstract description 51
- QAIPRVGONGVQAS-UHFFFAOYSA-N cis-caffeic acid Natural products OC(=O)C=CC1=CC=C(O)C(O)=C1 QAIPRVGONGVQAS-UHFFFAOYSA-N 0.000 claims abstract description 51
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 20
- 239000011259 mixed solution Substances 0.000 claims abstract description 12
- 238000002156 mixing Methods 0.000 claims abstract description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 27
- 238000010438 heat treatment Methods 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 15
- 239000000843 powder Substances 0.000 claims description 13
- 238000000634 powder X-ray diffraction Methods 0.000 claims description 13
- 238000002425 crystallisation Methods 0.000 claims description 7
- 230000008025 crystallization Effects 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 7
- 238000010521 absorption reaction Methods 0.000 claims description 6
- 238000000862 absorption spectrum Methods 0.000 claims description 6
- 230000008569 process Effects 0.000 claims description 6
- 206010028980 Neoplasm Diseases 0.000 claims description 4
- 238000000227 grinding Methods 0.000 claims description 4
- 206010018338 Glioma Diseases 0.000 claims description 3
- 238000001938 differential scanning calorimetry curve Methods 0.000 claims description 3
- 239000002246 antineoplastic agent Substances 0.000 claims description 2
- 208000035250 cutaneous malignant susceptibility to 1 melanoma Diseases 0.000 claims description 2
- 239000012467 final product Substances 0.000 claims description 2
- 208000029824 high grade glioma Diseases 0.000 claims description 2
- 201000011614 malignant glioma Diseases 0.000 claims description 2
- 201000001441 melanoma Diseases 0.000 claims description 2
- 229940041181 antineoplastic drug Drugs 0.000 claims 1
- 230000005496 eutectics Effects 0.000 abstract description 13
- 239000002994 raw material Substances 0.000 abstract description 13
- 238000011161 development Methods 0.000 abstract description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 229940079593 drug Drugs 0.000 description 12
- 239000000243 solution Substances 0.000 description 11
- 238000002844 melting Methods 0.000 description 10
- 230000008018 melting Effects 0.000 description 10
- 239000000203 mixture Substances 0.000 description 9
- 238000001228 spectrum Methods 0.000 description 8
- 230000004580 weight loss Effects 0.000 description 8
- 239000002904 solvent Substances 0.000 description 6
- 238000000859 sublimation Methods 0.000 description 6
- 230000008022 sublimation Effects 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 238000002447 crystallographic data Methods 0.000 description 4
- 238000001035 drying Methods 0.000 description 4
- -1 methyl diazonium ions Chemical class 0.000 description 4
- 238000013112 stability test Methods 0.000 description 4
- 239000007858 starting material Substances 0.000 description 4
- 238000001291 vacuum drying Methods 0.000 description 4
- 238000002441 X-ray diffraction Methods 0.000 description 3
- 230000000259 anti-tumor effect Effects 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 239000000306 component Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000000547 structure data Methods 0.000 description 3
- QRZMXADUXZADTF-UHFFFAOYSA-N 4-aminoimidazole Chemical compound NC1=CNC=N1 QRZMXADUXZADTF-UHFFFAOYSA-N 0.000 description 2
- MVBPAIHFZZKRGD-UHFFFAOYSA-N MTIC Chemical compound CNN=NC=1NC=NC=1C(N)=O MVBPAIHFZZKRGD-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 235000013824 polyphenols Nutrition 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000005855 radiation Effects 0.000 description 2
- 238000007670 refining Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000002076 thermal analysis method Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- VZSRBBMJRBPUNF-UHFFFAOYSA-N 2-(2,3-dihydro-1H-inden-2-ylamino)-N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]pyrimidine-5-carboxamide Chemical class C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C(=O)NCCC(N1CC2=C(CC1)NN=N2)=O VZSRBBMJRBPUNF-UHFFFAOYSA-N 0.000 description 1
- 206010051779 Bone marrow toxicity Diseases 0.000 description 1
- 208000003174 Brain Neoplasms Diseases 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 230000007067 DNA methylation Effects 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 238000004566 IR spectroscopy Methods 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 238000005411 Van der Waals force Methods 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N acrylic acid group Chemical group C(C=C)(=O)O NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 229940100198 alkylating agent Drugs 0.000 description 1
- 239000002168 alkylating agent Substances 0.000 description 1
- 230000002152 alkylating effect Effects 0.000 description 1
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008499 blood brain barrier function Effects 0.000 description 1
- 210000001218 blood-brain barrier Anatomy 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 231100000366 bone marrow toxicity Toxicity 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 229940044683 chemotherapy drug Drugs 0.000 description 1
- 238000002288 cocrystallisation Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000000113 differential scanning calorimetry Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 229910002804 graphite Inorganic materials 0.000 description 1
- 239000010439 graphite Substances 0.000 description 1
- 229910052736 halogen Inorganic materials 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- 239000012299 nitrogen atmosphere Substances 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000000879 optical micrograph Methods 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 239000005426 pharmaceutical component Substances 0.000 description 1
- 238000011170 pharmaceutical development Methods 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 229940002612 prodrug Drugs 0.000 description 1
- 239000000651 prodrug Substances 0.000 description 1
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 238000004467 single crystal X-ray diffraction Methods 0.000 description 1
- 238000000373 single-crystal X-ray diffraction data Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000012916 structural analysis Methods 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
- C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
- C07D487/04—Ortho-condensed systems
-
- 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
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C59/00—Compounds having carboxyl groups bound to acyclic carbon atoms and containing any of the groups OH, O—metal, —CHO, keto, ether, groups, groups, or groups
- C07C59/40—Unsaturated compounds
- C07C59/42—Unsaturated compounds containing hydroxy or O-metal groups
- C07C59/52—Unsaturated compounds containing hydroxy or O-metal groups a hydroxy or O-metal group being bound to a carbon atom of a six-membered aromatic ring
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B2200/00—Indexing scheme relating to specific properties of organic compounds
- C07B2200/13—Crystalline forms, e.g. polymorphs
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention belongs to the technical field of pharmaceutical co-crystals, and particularly relates to a temozolomide-caffeic acid pharmaceutical co-crystal and a preparation method thereof. The molecular formula of temozolomide-caffeic acid drug co-crystal is 2C 6 H 6 N 6 O 2 ·C 9 H 8 O 4 ·0.5H 2 O, the crystal structure belongs to monoclinic system, the space group is P2 (1)/c, and the unit cell parameter is α＝90°，β＝113631 (5) °, γ=90°, unit cell volumeMinimum asymmetric unit number z=8 in the unit cell. Uniformly mixing temozolomide with caffeic acid and acetone, and volatilizing and crystallizing the obtained temozolomide-caffeic acid mixed solution to obtain temozolomide-caffeic acid medicine eutectic. The temozolomide-caffeic acid pharmaceutical co-crystal prepared by the invention can effectively improve the stability and the solubility of a single raw material medicine of temozolomide and caffeic acid. The temozolomide-caffeic acid medicine co-crystal prepared by the invention has simple preparation method and is easy for industrial development and production.
Description
Technical Field
The invention belongs to the technical field of pharmaceutical co-crystals, and particularly relates to a temozolomide-caffeic acid pharmaceutical co-crystal and a preparation method thereof.
Background
Temozolomide (TMZ) is an oral alkylating agent used to treat malignant gliomas, which is fully absorbed orally, has a bioavailability of nearly 100%, shows broad spectrum activity in murine tumor models and is able to penetrate the human blood brain barrier. The cytotoxic effect of temozolomide is due to its strong methylation of DNA bases. Under alkaline conditions, temozolomide can be rapidly broken to generate active methyl diazonium ions. Because the brain tumor has higher alkalinity than surrounding tissues, the activation of the medicine can relatively occur in a concentrated way at the tumor part, the anti-tumor effect is strong and has certain selectivity, the side effect spectrum is improved, the bone marrow toxicity is smaller, and the tolerance of patients is improved. However it has an oral biological halfShort aging period, poor stability and tabletting property and the like, and limits the clinical curative effect of the composition. TMZ is a prodrug that spontaneously hydrolyzes under physiological conditions to the active compound 5- (3-methyl-1-triazine) imidazole-4-hydroxyamine (MTIC). MTIC is further decomposed into 5-aminoimidazole 4-hydroxylamine (AIC) and methyl diazonium Cation (CH) 3 N 2 + ) The latter is an active alkylating substance that can exert antitumor effects by DNA methylation. Currently, TMZ is a first-line chemotherapeutic drug. However, clinical use of TMZ is limited due to rapid elimination after oral administration. In addition, TMZ presents stability problems because it degrades to AIC during storage and processing, which also reduces its effectiveness.
Caffeic Acid (CA) is a polyphenol compound containing phenolic and acrylic functional groups. CA is reported to have antioxidant, antibacterial, antiviral and anticancer properties. Therefore, CA has gained considerable interest in the scientific community. However, CA has low bioavailability due to its poor solubility in aqueous environments, which limits its use in the food and pharmaceutical industries.
Pharmaceutical co-crystals are crystals formed by combining active pharmaceutical ingredients and proper co-crystal reagents under the action of ammonia bonds or other non-covalent bonds (pi-pi conjugation, van der Waals forces, halogen bonds and the like) by applying crystal engineering, pharmaceutical science, supermolecular chemistry principles and self-assembly principles. The pharmaceutical co-crystal is used as a new pharmaceutical crystal form, and can introduce new components without affecting the internal structure of the medicine, thereby greatly improving the physicochemical properties of the medicine and improving the clinical efficacy. Not only can the stability of the drug be ensured, but also the stability of the drug can be greatly changed in other aspects such as melting point, solubility and the like.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a temozolomide-caffeic acid medicine co-crystal and a preparation method thereof, and compared with the bulk drugs, the temozolomide-caffeic acid co-crystal has good chemical stability, improves the solubility of caffeic acid while reducing the solubility of temozolomide, and plays a synergistic effect.
In order to solve the technical problems, the invention adopts the following technical scheme:
the invention discloses a temozolomide-caffeic acid medicine co-crystal, the molecular formula of which is 2C 6 H 6 N 6 O 2 ·C 9 H 8 O 4 ·0.5H 2 O, the crystal structure belongs to monoclinic system, the space group is P2 (1)/c, and the unit cell parameter isα=90°, β= 113.631 (5) °, γ=90°, unit cell volume +.>Minimum asymmetric unit number z=8 in unit cell, crystal density of 1.546g/cm 3 ；
The powder X-ray diffraction pattern measured by Cu/K alpha rays after grinding shows that the 2 theta angle expressed in degrees is 11.47 degrees, 13.21 degrees, 13.71 degrees, 14.89 degrees, 16.08 degrees, 26.29 degrees, 26.51 degrees, 26.9 degrees, 27.92 degrees and 28.7 degrees, and has characteristic diffraction peaks;
the infrared absorption spectrum of the temozolomide-caffeic acid medicine co-crystal measured by KBr tabletting is 3535cm -1 、3433cm -1 、3349cm -1 、3300cm -1 、1741cm -1 、1667cm -1 、1595cm -1 、1519cm -1 、1458cm -1 、1360cm -1 、1265cm -1 、1179cm -1 A characteristic absorption peak is arranged at the position;
the temozolomide-caffeic acid drug co-crystal has an exothermic peak at 179-201 ℃ and the peak value is 184.7 ℃ by using a DSC curve measured by a differential scanning calorimeter;
the temozolomide-caffeic acid medicine co-crystal is crystalline powder.
Wherein, the sources of water molecules in the temozolomide-caffeic acid medicine co-crystal are as follows: (1) Water in the environment during crystallization in the process of preparing temozolomide-caffeic acid medicine co-crystal; and/or (2) the temozolomide raw material or the caffeic acid raw material contains crystal water.
In some embodiments, the crystalline powder has a bulk density of 0.59 to 0.65g/cm 3 。
In some embodiments, the crystalline powder has a tap density of 1.01 to 1.08g/cm 3 。
Further, the invention discloses a preparation method of the temozolomide-caffeic acid medicine co-crystal, which comprises the steps of uniformly mixing temozolomide with caffeic acid and acetone to obtain a temozolomide-caffeic acid mixed solution; volatilizing and crystallizing the temozolomide-caffeic acid mixed solution to obtain the final product.
Wherein, the preparation method of the temozolomide-caffeic acid drug co-crystal is a solution method.
In some embodiments, the molar ratio of temozolomide to caffeic acid is 2:1, a step of; the mol volume ratio of the temozolomide to the acetone is 0.2-1 mmol:10mL.
In some embodiments, water bath heating and stirring are required in the process of uniformly mixing temozolomide with caffeic acid and acetone.
Wherein the water bath heating is carried out, the heating temperature is 80-90 ℃, and the heating time is 1-2 h; preferably, the water bath is heated, the heating temperature is 80 ℃, and the heating time is 2 hours.
Wherein the stirring speed is 400-600 rpm, preferably 500 rpm.
In some embodiments, the volatile crystallization is performed at a crystallization temperature of 20 to 40 ℃, preferably 30 ℃, and a crystallization time of 1 to 2 weeks, preferably 1 week.
Wherein, the temozolomide-caffeic acid medicine co-crystal prepared by the preparation method is yellow and transparent.
And placing the crystal obtained after the mixed solution of temozolomide and caffeic acid is volatilized and crystallized in a vacuum drying oven, drying for 8 hours at 40 ℃, and volatilizing the solvent to obtain temozolomide-caffeic acid medicine eutectic crystal powder.
The application of the temozolomide-caffeic acid medicine co-crystal in preparing anti-tumor medicines is also within the protection scope of the invention.
In particular, the tumor is a malignant glioma or a malignant melanoma.
The beneficial effects are that:
(1) The temozolomide-caffeic acid pharmaceutical co-crystal prepared by the invention can effectively improve the stability and the solubility of a single raw material medicine of temozolomide and caffeic acid.
(2) The temozolomide-caffeic acid medicine co-crystal prepared by the invention provides a better medication strategy for combined medication.
(3) The temozolomide-caffeic acid medicine co-crystal prepared by the invention has simple preparation method and is easy for industrial development and production.
Drawings
The foregoing and/or other advantages of the invention will become more apparent from the following detailed description of the invention when taken in conjunction with the accompanying drawings and detailed description.
FIG. 1 is a unit cell packing diagram of temozolomide-caffeic acid co-crystal in example 1 of the present invention.
FIG. 2 is an X-ray diffraction pattern of temozolomide-caffeic acid co-crystal in example 1 of the present invention.
FIG. 3 is an X-ray diffraction pattern of temozolomide starting material used in the examples of the present invention.
FIG. 4 is an X-ray diffraction pattern of caffeic acid materials used in examples of the invention.
FIG. 5 is a graph of Differential Scanning Calorimetric (DSC) analysis of temozolomide, caffeic acid, and temozolomide-caffeic acid co-crystals in example 1 of the present invention.
FIG. 6 is a Thermogravimetric (TGA) analysis of temozolomide, caffeic acid and temozolomide-caffeic acid co-crystals in example 1 of the present invention.
FIG. 7 is an infrared spectrum of temozolomide, caffeic acid and temozolomide-caffeic acid co-crystal in example 1 of the present invention.
FIG. 8 is a graph showing the dissolution of temozolomide, caffeic acid and temozolomide-caffeic acid co-crystals in pure water in example 1 of the present invention.
Fig. 9 is an optical microscope photograph of temozolomide-caffeic acid co-crystal in example 1 of the present invention.
FIG. 10 is a schematic representation of temozolomide starting material in example 1 of the present invention 1 H NMR chart.
FIG. 11 shows the caffeic acid feed in example 1 of the invention 1 H NMR chart.
FIG. 12 is a schematic diagram of temozolomide-caffeic acid co-crystal in example 1 of the invention 1 H NMR chart.
FIG. 13 is a graph showing the accelerated stability test of temozolomide-caffeic acid co-crystal in example 1 of the present invention.
Detailed Description
For further explanation of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative efforts fall within the protection scope of the present invention.
1. Reagent(s)
The temozolomide raw material used in the embodiment of the invention is purchased from Shanghai microphone Biochemical technology Co., ltd, the purity is 99%, and the powder X-ray diffraction spectrum of the temozolomide raw material is shown in figure 3.
The caffeic acid material used in the examples of the present invention was purchased from Shanghai Mai Rui Biochemical technology Co., ltd, and the purity was 99%, and the powder X-ray diffraction pattern of the caffeic acid material was shown in FIG. 4.
2. Measurement method
Single crystal X-ray diffraction determination of crystal structure: the single crystal thus grown was cut into a size of about 0.10X0.20X0.30 mm 3 Block of the size, measured by a BrookAPEX-IICCD diffractometerMo/ka radiation source (graphite monochromator,) And irradiating the sample, collecting diffraction data, reducing the diffraction data by SAINT, performing structural analysis by using a SHELXTL software direct method, and refining all non-hydrogen atoms by using an anisotropic refining method based on a full matrix least square method of F2.
Powder X-ray diffraction: the ground sample was taken at about 0.1g and diffraction data was collected by a powder X-ray diffractometer (Bruker D8 Advance) at room temperature with Cu/K alpha radiation as the light sourceThe scanning step length is 0.02 degrees, the scanning voltage is set to be 40kV, the current is set to be 40mA, the scanning speed is set to be 0.2s/0.02 degrees, the scanning range 2 theta is set to be 5-40 degrees, and data are processed through Jade software and plotted through Origin.
Fourier infrared spectroscopy: mixing 20mg of crystalline powder with 400mg of dry KBr, grinding, pressing into slices by using a tablet press, and collecting infrared absorption spectrum data at room temperature by using a Fourier infrared spectrometer (Nigao instruments/Nanjin Zhonghui scientific equipment Co., ltd.), wherein the spectrum range is as follows: 4000-400 cm -1 Resolution: 0.1, linearity: 0.1% t.
Thermal analysis: each sample was placed on an alumina crucible using simultaneous thermal analysis (manufactured by the company of nai instruments, germany/the company of sciences equipment, south kyo new), heated from 40 ℃ to 350 ℃ at a heating rate of 10 ℃/min in a nitrogen atmosphere, and thermal weight and differential thermal data information were obtained simultaneously with the same sample in the same measurement.
Nuclear magnetic measurement: weighing 10-30 mg of sample, taking dimethyl sulfoxide-d 6 as solvent, taking TMS (0 ppm) as internal standard, and measuring on a AVANCE III MHz Bruker digital spectrometer to obtain 1 H NMR spectrum.
The method for measuring the solubility comprises the following steps: respectively weighing excessive temozolomide, caffeic acid and temozolomide-caffeic acid medicine eutectic, respectively putting into 20mL of pure water, dissolving in water bath at the speed of 100rpm and the temperature of 37 ℃, respectively taking 1mL of solution at the 5 th, 15 th, 30 th, 60 th, 90 th, 120 th, 180 th and 250 th min, simultaneously supplementing the medium with the same volume, diluting the taken solution by a proper multiple after passing through a water system microporous filter membrane, and measuring by using an island body fluid UV-3600 ultraviolet spectrophotometer.
Acceleration stability test: 100mg of TMZ-CA eutectic crystal powder is weighed and evenly spread into an open culture dish, the thickness is less than or equal to 5mm, the mixture is placed into a constant temperature incubator with the temperature of 40 ℃ and the relative humidity of 75+/-5%, and the mixture is sampled and detected in 1 st month, 2 nd month and 3 rd month respectively, and is compared with the result of 0 day.
Example 1
Preparation of co-crystals: mixing 1mmol of temozolomide and 0.5mmol of caffeic acid, placing the mixture into a 20mL glass bottle, adding 10mL of acetone solution, heating in a water bath to 80 ℃, stirring at a speed of 500r/min for 2h, and heating in the water bath to dissolve the mixture to obtain transparent liquid, thus obtaining the temozolomide-caffeic acid mixed solution. Filtering the temozolomide-caffeic acid mixed solution to a 20mL small bottle through a 0.22 mu m organic filter head, standing the solution in an incubator at 30 ℃ for volatilizing and crystallizing for 1 week to obtain a yellow transparent temozolomide-caffeic acid medicine eutectic. Placing the obtained drug co-crystal into a vacuum drying oven, drying at 40deg.C for 8 hr, volatilizing solvent to obtain temozolomide-caffeic acid drug co-crystal powder with 90% yield and 0.65g/cm bulk density 3 Tap density of 1.08g/cm 3 。
Characterization of temozolomide-caffeic acid drug co-crystal:
(1) The obtained temozolomide-caffeic acid medicine co-crystal is subjected to X-ray single crystal diffraction, the crystal structure data are shown in figure 1, and as can be seen from figure 1, the temozolomide-caffeic acid medicine co-crystal has a channel type structure, and the structure comprises 2 layers, namely one layer contains water and the other layer does not contain water.
The molecular formula of the obtained temozolomide-caffeic acid drug co-crystal is 2C 6 H 6 N 6 O 2 ·C 9 H 8 O 4 ·0.5H 2 O, the crystal structure belongs to monoclinic system, the space group is P2 (1)/c, and the unit cell parameter is α=90°, β= 113.631 (5) °, γ=90°, unit cell volume +.>Minimum asymmetric unit number z=8 in unit cell, crystal density of 1.546g/cm 3 Crystal size 0.4X0.36 X0.25 mm 3 Specific single crystal X-ray diffraction data are shown in table 1.
TABLE 1 single crystal X-diffraction data for temozolomide-caffeic acid drug co-crystals
(2) The temozolomide-caffeic acid medicine co-crystal is ground to carry out X-ray powder diffraction, the powder X-ray diffraction spectrum is shown in figure 2, and the powder X-ray diffraction spectrum measured by Cu/K alpha rays after grinding the temozolomide-caffeic acid medicine co-crystal has characteristic diffraction peaks at the positions of 11.47 degrees, 13.21 degrees, 13.71 degrees, 14.89 degrees, 16.08 degrees, 26.29 degrees, 26.51 degrees, 26.9 degrees, 27.92 degrees and 28.7 degrees. The obtained temozolomide-caffeic acid co-crystal has obvious difference compared with temozolomide and caffeic acid raw materials, wherein powder X-ray diffraction patterns of the temozolomide and the caffeic acid raw materials are respectively shown in figure 3 and figure 4.
(3) The DSC curve of temozolomide-caffeic acid drug co-crystal measured using a differential scanning calorimeter is shown in fig. 5, from which it can be seen by differential scanning calorimeter analysis chart: (i) The temozolomide raw material has an exothermic peak in the heating process, the peak temperature is 205.3 ℃, no phase change peak is seen in the cooling process, the exothermic peak of a single crystal form is displayed, and the melting point of the crystal is 205.3 ℃; (ii) The raw material caffeic acid has an endothermic peak and an exothermic peak in the heating process, the peak temperatures are 234.2 ℃ and 241.3 ℃ respectively, and no phase change peak is seen in the cooling process; (iii) The temozolomide-caffeic acid eutectic has an exothermic peak in the heating process, and the peak temperature is 184.7 ℃, which shows that the melting point is 184.7 ℃ and is lower than the melting points of temozolomide and caffeic acid serving as raw materials.
(4) The thermal weight curve of temozolomide-caffeic acid drug co-crystal is shown in fig. 6, and it can be seen from the TG curve that neither TMZ nor CA has a weight loss plateau, indicating that neither sample contains crystal water or other solvents, and starts to decompose at 191.26 ℃ and 196.90 ℃, respectively. As can be seen from the TG curve, the water loss phenomenon starts to appear when TMZ-CA (temozolomide-caffeic acid drug co-crystal) is heated to 100 ℃, which indicates that the sample contains crystal water, the weight loss of the sample is 1.6%, and the content of 1/2 water molecules in the corresponding single crystal structure is consistent with that of the single crystal structure.
Before the temozolomide reaches the melting point (205.3 ℃), sublimation occurs to cause weight loss to a certain extent, and the weight loss is accelerated until complete sublimation after the temozolomide reaches the melting point; the caffeic acid undergoes sublimation before the melting point (234.2 ℃) to cause weight loss to a certain extent, and the weight loss is accelerated until complete sublimation after reaching the melting point; the temozolomide-caffeic acid co-crystal undergoes sublimation before the melting point (184.7 ℃) to cause a certain degree of weight loss, and the weight loss is accelerated until complete sublimation after reaching the melting point.
(5) At 4000-400 cm -1 The infrared absorption spectrum of the temozolomide, caffeic acid and temozolomide-caffeic acid co-crystal is shown in figure 7, wherein the infrared absorption spectrum of the temozolomide-caffeic acid drug co-crystal is 3535cm -1 、3433cm -1 、3349cm -1 、3300cm -1 、1741cm -1 、1667cm -1 、1595cm -1 、1519cm -1 、1458cm -1 、1360cm -1 、1265cm -1 、1179cm -1 There is a characteristic absorption peak.
It can be noted that in the spectrum of the temozolomide-caffeic acid co-crystal, a part of the absorption peak is shifted and changed in peak shape and at 3535cm, compared with the infrared absorption spectrum of the raw material temozolomide, caffeic acid -1 Where a new feature absorption occurs,indicating that temozolomide and caffeic acid molecules are involved in the formation of new hydrogen bond interactions, indicating the formation of temozolomide-caffeic acid co-crystals. Notably, in the infrared spectrum of the co-crystal, the shift in the absorption band means that caffeic acid is involved in co-crystallization with temozolomide and indicates the formation of the co-crystal.
(6) Fig. 8 is a graph showing the dissolution profiles of temozolomide, caffeic acid and temozolomide-caffeic acid co-crystals in pure water, wherein the temozolomide-caffeic acid drug co-crystals have higher solubility in pure water solution (CA vs CA in the co-crystal) than the drug caffeic acid and lower solubility in water (TMZ vs TMZ in the co-crystal) than the drug temozolomide.
(7) An optical microscope image of the temozolomide-caffeic acid co-crystal prepared in this example 1 is shown in fig. 9.
(8) Temozolomide starting material, caffeic acid starting material and temozolomide-caffeic acid co-crystal prepared in this example 1 1 HNMR spectra are shown in fig. 10, 11, and 12, respectively.
By using 1 H NMR was measured to determine the stoichiometry and chemical purity of each eutectic form. The obtained eutectic crystal 1 The H NMR spectrum is the sum of the characteristic peaks of TMZ and CA, indicating that these two components are present in the new phase. Of eutectic products 1 The H NMR chemical shift distribution is as follows: 1 H NMR(400MHz,DMSO-d6)δ12.12(s,1H),9.48(s,1H),9.18(s,1H),8.82(s,2H),7.79(s,2H),7.67(s,2H),7.41(d,J＝15.9Hz,1H),7.02(d,J＝2.1Hz,1H),6.96(dd,J＝8.2,2.1Hz,1H),6.76(d,J＝8.1Hz,1H),6.17(d,J＝15.9Hz,1H),3.87(s,6H)。
TMZ and CA in Co-crystallized product 1 The H NMR chemical shift distribution is as follows: TMZ 1 H NMR (400 MHz, DMSO-d 6) delta 8.82 (s, 1H), 7.79 (s, 1H), 7.67 (s, 1H), 3.87 (s, 3H); CA (CA) 1 H NMR (400 mhz, dmso-d 6) δ12.11 (s, 1H), 9.52 (s, 1H), 9.14 (s, 1H), 7.42 (d, j=15.9 hz, 1H), 7.03 (d, j=2.1 hz, 1H), 6.97 (dd, j=8.2, 2.1hz, 1H), 6.76 (d, j=8.1 hz, 1H), 6.18 (d, j=15.9 hz, 1H). From the integral of the single characteristic proton signal, the stoichiometric ratio of TMZ and CA was calculated as 2:1, a step of; at the same time, the nuclear magnetic hydrogen spectrum peak yield of the eutecticA slight shift is generated, which may be due to eutectic formation.
(9) The accelerated stability test of temozolomide-caffeic acid co-crystal prepared in this example 1 is shown in fig. 13. Stability tests show that compared with temozolomide which is spontaneously hydrolyzed and decomposed into 5-aminoimidazole 4-hydroxylamine (AIC) after three months, the temozolomide-caffeic acid eutectic has no crystal phase change within three months, and has remarkable physical and chemical stability.
Example 2: preparation of temozolomide-caffeic acid co-crystal
Mixing 1mmol of temozolomide and 0.5mmol of caffeic acid, placing the mixture into a 20mL glass bottle, adding 10mL of acetone solution, heating in a water bath to 90 ℃, stirring at a speed of 500r/min for 1h, and heating in the water bath to dissolve the mixture to obtain transparent liquid, thus obtaining the temozolomide-caffeic acid mixed solution. Filtering the temozolomide-caffeic acid mixed solution to a 20mL small bottle through a 0.22 mu m organic filter head, standing the solution in an incubator at 30 ℃ for volatilizing and crystallizing for 1 week to obtain a yellow transparent temozolomide-caffeic acid medicine eutectic. Placing the obtained drug co-crystal into a vacuum drying oven, drying at 40deg.C for 8 hr, volatilizing solvent to obtain temozolomide-caffeic acid drug co-crystal powder with yield of 87% and bulk density of 0.62g/cm 3 Tap density of 1.03g/cm 3 。
The obtained co-crystal was subjected to X-ray single crystal diffraction, and the co-crystal structure data was identical to that of the co-crystal obtained in example 1. The co-crystal was ground and subjected to X-ray powder diffraction, and the powder X-ray diffraction pattern was identical to that of the co-crystal obtained in example 1.
Example 3: preparation of temozolomide-caffeic acid co-crystal
Mixing 1mmol of temozolomide and 0.5mmol of caffeic acid, placing the mixture into a 20mL glass bottle, adding 10mL of acetone solution, heating in a water bath to 90 ℃, stirring at a speed of 500r/min for 2h, and heating in the water bath to dissolve the mixture to obtain transparent liquid, thus obtaining the temozolomide-caffeic acid mixed solution. Filtering temozolomide-caffeic acid mixed solution to 20mL vial through 0.22 μm organic filter head, and addingAnd (3) in a constant temperature cabinet at 30 ℃, standing the solution for volatilizing and crystallizing for 1 week to obtain a yellow transparent temozolomide-caffeic acid medicine eutectic. Placing the obtained drug co-crystal into a vacuum drying oven, drying at 40deg.C for 8 hr, volatilizing solvent to obtain temozolomide-caffeic acid drug co-crystal powder with 89% yield and 0.59g/cm bulk density 3 Tap density of 1.01g/cm 3 。
The obtained crystal was subjected to X-ray single crystal diffraction, and the crystal structure data were identical to those of the crystal obtained in example 1. The crystals were ground and subjected to X-ray powder diffraction, the powder X-ray diffraction pattern being in accordance with example 1.
In conclusion, the invention obtains a novel drug co-crystal, namely temozolomide-caffeic acid co-crystal through research. The temozolomide-caffeic acid medicine co-crystal provided by the invention improves the solubility of raw material caffeic acid and reduces the solubility of temozolomide. The temozolomide-caffeic acid pharmaceutical co-crystal improves the solubility, stability and other physicochemical properties of the pharmaceutical components on the premise of not changing the molecular structures of temozolomide and caffeic acid pharmaceutical, and the pharmaceutical co-crystal is widely accepted by the pharmaceutical industry along with the occurrence of more and more pharmaceutical co-crystal researches, so that the temozolomide-caffeic acid pharmaceutical co-crystal becomes an important ring of pharmaceutical development work.
The invention provides a temozolomide-caffeic acid pharmaceutical co-crystal, a method for preparing the same, and a method for realizing the technical scheme. The components not explicitly described in this embodiment can be implemented by using the prior art.
Claims (10)
1. A temozolomide-caffeic acid medicine co-crystal is characterized in that the molecular formula of the temozolomide-caffeic acid medicine co-crystal is 2C 6 H 6 N 6 O 2 ·C 9 H 8 O 4 ·0.5H 2 O, the crystal structure belongs to monoclinic system, the space group is P2 (1)/c, and the unit cell parameter isα=90°, β= 113.631 (5) °, γ=90°, unit cell volume +.>Minimum asymmetric unit number z=8 in unit cell, crystal density of 1.546g/cm 3 ；
The powder X-ray diffraction pattern measured by Cu/K alpha rays after grinding shows that the 2 theta angle expressed in degrees is 11.47 degrees, 13.21 degrees, 13.71 degrees, 14.89 degrees, 16.08 degrees, 26.29 degrees, 26.51 degrees, 26.9 degrees, 27.92 degrees and 28.7 degrees, and has characteristic diffraction peaks;
the infrared absorption spectrum of the temozolomide-caffeic acid medicine co-crystal measured by KBr tabletting is 3535cm -1 、3433cm -1 、3349cm -1 、3300cm -1 、1741cm -1 、1667cm -1 、1595cm -1 、1519cm -1 、1458cm -1 、1360cm -1 、1265cm -1 、1179cm -1 A characteristic absorption peak is arranged at the position;
the temozolomide-caffeic acid drug co-crystal has an exothermic peak at 179-201 ℃ and the peak value is 184.7 ℃ by using a DSC curve measured by a differential scanning calorimeter;
the temozolomide-caffeic acid medicine co-crystal is crystalline powder.
2. The moxazolamide-caffeic acid pharmaceutical co-crystal according to claim 1, wherein the bulk density of the crystalline powder is 0.59-0.65 g/cm 3 。
3. The moxazolamide-caffeic acid pharmaceutical co-crystal according to claim 1, wherein the crystalline powder has a tap density of 1.01-1.08 g/cm 3 。
4. The method for preparing temozolomide-caffeic acid pharmaceutical co-crystal according to claim 1, which is characterized in that temozolomide is uniformly mixed with caffeic acid and acetone to obtain temozolomide-caffeic acid mixed solution; volatilizing and crystallizing the temozolomide-caffeic acid mixed solution to obtain the final product.
5. The method according to claim 4, wherein the molar ratio of temozolomide to caffeic acid is 2:1, a step of; the mol volume ratio of the temozolomide to the acetone is 0.2-1 mmol:10mL.
6. The preparation method according to claim 4, wherein water bath heating and stirring are required in the process of uniformly mixing temozolomide with caffeic acid and acetone.
7. The preparation method according to claim 6, wherein the water bath heating is performed at 80-90 ℃ for 1-2 hours; the stirring speed is 400-600 rpm, preferably 500 rpm.
8. The process according to claim 4, wherein the volatile crystallization is carried out at a crystallization temperature of 20 to 40 ℃, preferably 30 ℃ for a crystallization time of 1 to 2 weeks.
9. Use of temozolomide-caffeic acid pharmaceutical co-crystal according to claim 1 in the preparation of an antitumor drug.
10. The use according to claim 9, wherein the tumour is a malignant glioma or a malignant melanoma.
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