CA2097611A1 - Verbenone, having antielastase actibity, against pulmonary emphysema - Google Patents
Verbenone, having antielastase actibity, against pulmonary emphysemaInfo
- Publication number
- CA2097611A1 CA2097611A1 CA002097611A CA2097611A CA2097611A1 CA 2097611 A1 CA2097611 A1 CA 2097611A1 CA 002097611 A CA002097611 A CA 002097611A CA 2097611 A CA2097611 A CA 2097611A CA 2097611 A1 CA2097611 A1 CA 2097611A1
- Authority
- CA
- Canada
- Prior art keywords
- verbenone
- pulmonary emphysema
- antielastase
- activity
- actibity
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 206010014561 Emphysema Diseases 0.000 title claims abstract description 11
- 102100022712 Alpha-1-antitrypsin Human genes 0.000 title claims description 5
- 101000823116 Homo sapiens Alpha-1-antitrypsin Proteins 0.000 title claims description 5
- 101001010513 Homo sapiens Leukocyte elastase inhibitor Proteins 0.000 title claims description 5
- DCSCXTJOXBUFGB-JGVFFNPUSA-N (R)-(+)-Verbenone Natural products CC1=CC(=O)[C@@H]2C(C)(C)[C@H]1C2 DCSCXTJOXBUFGB-JGVFFNPUSA-N 0.000 title description 22
- DCSCXTJOXBUFGB-SFYZADRCSA-N (R)-(+)-verbenone Chemical compound CC1=CC(=O)[C@H]2C(C)(C)[C@@H]1C2 DCSCXTJOXBUFGB-SFYZADRCSA-N 0.000 title description 20
- DCSCXTJOXBUFGB-UHFFFAOYSA-N verbenone Natural products CC1=CC(=O)C2C(C)(C)C1C2 DCSCXTJOXBUFGB-UHFFFAOYSA-N 0.000 title description 20
- 239000003814 drug Substances 0.000 claims abstract description 11
- 238000002360 preparation method Methods 0.000 claims abstract description 7
- 238000011282 treatment Methods 0.000 claims abstract description 7
- 230000000694 effects Effects 0.000 claims description 13
- 239000000243 solution Substances 0.000 claims description 10
- 239000000203 mixture Substances 0.000 claims description 8
- 229920000858 Cyclodextrin Polymers 0.000 claims description 7
- 239000000443 aerosol Substances 0.000 claims description 3
- 239000008187 granular material Substances 0.000 claims description 3
- 239000007923 nasal drop Substances 0.000 claims description 3
- 229940100662 nasal drops Drugs 0.000 claims description 3
- 239000006188 syrup Substances 0.000 claims description 3
- 235000020357 syrup Nutrition 0.000 claims description 3
- 239000002775 capsule Substances 0.000 claims description 2
- 239000008298 dragée Substances 0.000 claims description 2
- 238000007918 intramuscular administration Methods 0.000 claims description 2
- 238000001990 intravenous administration Methods 0.000 claims description 2
- 239000008194 pharmaceutical composition Substances 0.000 claims description 2
- 239000006187 pill Substances 0.000 claims description 2
- 239000000829 suppository Substances 0.000 claims description 2
- 239000003826 tablet Substances 0.000 claims description 2
- 239000000725 suspension Substances 0.000 claims 1
- 150000003505 terpenes Chemical class 0.000 abstract description 2
- 235000007586 terpenes Nutrition 0.000 abstract description 2
- 102000016387 Pancreatic elastase Human genes 0.000 description 8
- 108010067372 Pancreatic elastase Proteins 0.000 description 8
- 102000016942 Elastin Human genes 0.000 description 6
- 108010014258 Elastin Proteins 0.000 description 6
- 108091005804 Peptidases Proteins 0.000 description 6
- 239000004365 Protease Substances 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- 229920002549 elastin Polymers 0.000 description 6
- 230000003448 neutrophilic effect Effects 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 230000000510 mucolytic effect Effects 0.000 description 4
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 102000035195 Peptidases Human genes 0.000 description 3
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 3
- 230000003110 anti-inflammatory effect Effects 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- MUMGGOZAMZWBJJ-DYKIIFRCSA-N Testostosterone Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 MUMGGOZAMZWBJJ-DYKIIFRCSA-N 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- WHGYBXFWUBPSRW-FOUAGVGXSA-N beta-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 210000003456 pulmonary alveoli Anatomy 0.000 description 2
- 210000002345 respiratory system Anatomy 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 101710081722 Antitrypsin Proteins 0.000 description 1
- 208000017667 Chronic Disease Diseases 0.000 description 1
- 206010010356 Congenital anomaly Diseases 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 208000019693 Lung disease Diseases 0.000 description 1
- 241000208125 Nicotiana Species 0.000 description 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- GBOGMAARMMDZGR-UHFFFAOYSA-N UNPD149280 Natural products N1C(=O)C23OC(=O)C=CC(O)CCCC(C)CC=CC3C(O)C(=C)C(C)C2C1CC1=CC=CC=C1 GBOGMAARMMDZGR-UHFFFAOYSA-N 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 230000001475 anti-trypsic effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
- 210000003123 bronchiole Anatomy 0.000 description 1
- 206010006451 bronchitis Diseases 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- GBOGMAARMMDZGR-JREHFAHYSA-N cytochalasin B Natural products C[C@H]1CCC[C@@H](O)C=CC(=O)O[C@@]23[C@H](C=CC1)[C@H](O)C(=C)[C@@H](C)[C@@H]2[C@H](Cc4ccccc4)NC3=O GBOGMAARMMDZGR-JREHFAHYSA-N 0.000 description 1
- GBOGMAARMMDZGR-TYHYBEHESA-N cytochalasin B Chemical compound C([C@H]1[C@@H]2[C@@H](C([C@@H](O)[C@@H]3/C=C/C[C@H](C)CCC[C@@H](O)/C=C/C(=O)O[C@@]23C(=O)N1)=C)C)C1=CC=CC=C1 GBOGMAARMMDZGR-TYHYBEHESA-N 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- POZRVZJJTULAOH-LHZXLZLDSA-N danazol Chemical compound C1[C@]2(C)[C@H]3CC[C@](C)([C@](CC4)(O)C#C)[C@@H]4[C@@H]3CCC2=CC2=C1C=NO2 POZRVZJJTULAOH-LHZXLZLDSA-N 0.000 description 1
- 229960000766 danazol Drugs 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000001524 infective effect Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 150000002773 monoterpene derivatives Chemical class 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 210000003097 mucus Anatomy 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 210000001539 phagocyte Anatomy 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 208000023504 respiratory system disease Diseases 0.000 description 1
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000779 smoke Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 229960003604 testosterone Drugs 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000002753 trypsin inhibitor Substances 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
-
- 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
- A61K31/12—Ketones
-
- 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
- A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6949—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
- A61K47/6951—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes using cyclodextrin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nanotechnology (AREA)
- Epidemiology (AREA)
- Medical Informatics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Pulmonology (AREA)
- Medicinal Preparation (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Saccharide Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The use of a terpene derivative for the preparation of a medicament useful in the treatment of pulmonary emphysema.
Description
w o 93/06823 2 0 9 7 611 PCT/EP92/02269 VFRBENONE, HAVING ANTIELASTASE ACTIVITY,AGAINST PUUMONARY EMPHYSEMA
The present invention relates to the use of a terpene derivative, namely (lS)(-)-4,6,6-trimethylbicyclo-(3,1,1)-hept-3-ene-2-one (hereinafter named verbenone) of formula S O
~ 3 C~
~3 for the preparation of a medicament useful in the treatment of pulmonary emphysema. The invention also relates to a verbenone-cyclodextrin complex and the pharmaceutical compositions containing i~.
Pulmonary emphysema is a progressive respiratory disorder characterized by an abnormal and permanent increase in the volume of the air spaces located distally to the terminal bronchiole, with destruction of the alveolar septum.
A number of experimental and clinical data generally ascribe the emphysema pathogenesis to an unbalance of the protease-antiprotease ratio at the alveolus level. Under normal conditions, alveolar proteases exert an essential activity, which is the lysis of cell fragments, thrombi and particles. Said activity is counterbalanced be endogenous antiproteases ~ 1 antitrypsin [a-lAT], ~-2 macroglob~lin [~-2M], .. ~,,~ . .
.
Bronchial Mucus Inhibitor [B.M.I.]), which yrotect the Dulmonary tissue from the protease action. The loss of the normal balance between proteases and antlproteases can be attributed to three factors: 1) congenital lack in antiproteases (particularly ~-lAT); 2) excessive protease release at the alveolus level; 3) partial inactivation of antiproteases (~-lAT1 in the presence of a normal or enhanced protease production. Said factors are generally worsened by tobacco smoke.
Among alveolar proteases, leukocytal elastase proves to have the most marked digestive activity on all the pulmonary connective components, particularly on elastin, the fibres of which constitute the alveolar septa, thus giving lung the functional elasticity.
Besides the traditional treatments, which cannot remedy the elastin destruction, therapies have been studied to restore elastin synthesis and to protect elastin from elastases action. Satisfactory experimental results were obtained both administering human a-lAT intravenously and stimulating the hepatic synthesis thereof by administration of danatrol, which is a testosterone semi-synthetic derivative. However, both methods suffers from drawbacks, the first method being particularly complex and expensive, whereas the ~econd has antigonadotropinic effects.
Therefore there i5 the need for a medicament exerting an antielastase activity, directly or indi-rectly, and also having poor side-effects.
Now it has been found that verbenone exerts an unpredictable antielestase activity at the lung level, and, dl~e to said activity, it is use~ul in the - ;- .. .. .. .
treatment of emphysema.
Verbenone is a monoterpene derivative havins antiinflammatory and mucolytic activities and it is used as a mucolytic and fluidifying agent in the acute and chronic disorders of the respiratory tract.
As a consequence of further researches, verbenone showed a surprising inhibiting activity on the elastase action, particularly on the neutrophylic one. Said activity was unforeseeable from the pharmacological and clinical studies which evidenced the mucolytic and antiinflammatory activities of verbenone and the use thereof in therapy.
The use of verbenone in the treatment of pulmonary emphysema is extremely advantageous. In fact, pulmonary emphysema can be favoured, or worsened, by the simultaneous presence of other pulmonary disorders, such as bronchitis and/or pneumonia, other infective forms, asthma, which involve obstructions of the respiratory tract; therefore a medicament which can at the same time have antiinflammatory and mucolytic activities and protect elastin of the alveolar septa is obviously useful.
Tests were carried out in vitro to evaluate verbenone activity on the release of neutrophilic elastin from polymorphonuclear granules. Said tests evidenced the marked antielastase activity of verbenone, which activity is not dose-related nor cyto~oxic.
The results from said tests are reported hereinbelow.
Separation of neutro~hilic polymorphonuclears Polymorphonuclear phagocytes were separated starting from heparinized peripheral blood (60 cc) from healthy volunteers or from buffy coats (leukocytes and platelets enrichments).
Neutrophilic polymorphonuclears were separated by means of a 6~ colloidal dextran gradient, then the erythrocytes were lysated with an ammonium chloride solution.
Neutro hilic elastase release p 100 ,ul of cells at a concentration of lxlO7/ml, pretreated with cytochalasin B (5 ~g/ml) for 10', were incubated or 20' at 37C in a microtiter plate in the presence or in the absence of 50 yl of a challenge consisting ~f fMLP (formyl-methionyl-leuc~l-peptide) at a 10-6 molar inal concentration. After said incubation, the plate was centrifuged and 100 ~1 of supernatant were collected and incubated for 60-120' at 37C with a specific substrate for the neutrophilic elastase (methoxysuccinyl-ala-ala-pro-val-p-Na) to a 1 mM final concentration. The reaction was stopped by adding 50 ~1 of 1 N acetic acid and reading was carried out by means of a multiscan photometer at a 405 nm wave length. The values of the absorbance of samples challenged with f.~LP were compared to those from unchallenged cells, to obtain a ~ value corresponding to the amount of neutrophilic elastase released following to challenge. Any interferences of verbenone on the neutrophilic elastase release were tested by addition of different concentrations of the medicament directly to the cell suspension during the test.
,.. ,. . ; . . . . : .
' ' ' ........ ' ' ' ~ ............. . '' ,:
: : -. ,. . .
: . ~ ' . : : ' ' The results are shown in Table 1.
Modulation of the elastase release fro~ neutrophilic polymorphonuclears by addition of verbenone to the cells during the test, at different concentrations.
-% inhibitioncompared to the control Control Verb. 1 mM 90%
Verb. 0,1 mM 86%
Verb. 10 uM 94%
Verb. 1 pM 94%
Verb. 0,1 yM 92%
Verb. 10 nM 89%
Verb. 1 nM 83%
Verb. 0,1 nM 96%
Verb. 10 pM 94%
This marked inhibiting activity is unlikely to be ascribed to a toxic action of the medicament on the cells. In fact the cell viability, as repeatedly evaluated by the tripan bleu exclusion test, turned out to be always higher than 90% after incubation of the cells with verbenone in concentrations varying from 1 mM to lQ pM.
From the industrial point of view, verbenone is used as the active ingredient for the preparation of a medicament useful for the treatment of pulmonary emphysema.
::; . . .. ';` ' ' : .
For the preparation medicaments, verbenone has the drawbacks to be insoluble in water and to have a very unpleasant taste, therefore it cannot be used for aqueous liquid pharmaceutical forms, such as syrups, S oral solutions, nasal drops, aqueous solutions for aerosol and nebulizat on.
It has been found that the verbenone-cyclodextrin complex has high water solubility and, moreover, verbenone unpleasant taste is no more perceptible.
Therefore, the verbenone-cyclodextrin complex is a further object of invention. A further advantage of the complex of the invention is provided by its increased bioavailability.
The pharmaceutical forms comprising the verbenone-cyclodextrin complex can be prepared, for example,according to the methods described in "Remington's Pharmaceutical Sciences Handboo~", Hack Pub. Co., U~A.
Examples of pharmaceutical forms are capsules, pills, tablets, sugar-coated pills, granulates, syrups, drinkable solutions, nasal drops, solutions for aerosol or inhalation, suppositories, injectable solutions or sus~ension~ both for the intravenous and the intramuscular routes.
The dosages will depend on the severity of the disorder, the age, weight and sex of the patient and on the clinician's advise. Examples of dosages comprise single or repeated administrations of the medicament containing verbenone or the verbenone-cyclodextrin complex so as to attain a daily dosage from 10 to 500 mg of the active ingredient. The preparation of the verbenone-cyclod_xtrin complex is illustrated in the .... . .. . . ......... . . . .
~.
:' ' : . . .
: ~ .
W093/06823 2 0 9 7 ~11 PCT/EP92/02269 followin~ examples.
A ~-ciclodextrin amount equal to 0.1 mole is weighed and placed into a mortar of suitable size.
Verbenone ~0.1 mole), previously diluted with 15 ml of 96% ethanol, is added, and the mixture is mixed with a pestle to obtain an homogeneous mass. 20 ml of distilled water are added, and a mixture is obtained which hardens upon mixing. Said mixture is worked in a ~ortar for about 15 minutes, then it is dried in oven thermostatized at 60C.
The obtained complex is in form of a flowing powder which is completely odourless.
Verbenone titre ~ 80% on theoretical.
XA~PL~ 2 5.80 g o verbenone were dissolved in 100 ml of 95C ethanol (to obtain 50 g of a mixture), then they were added to 44.3 g of B-cyclodextrin in a round bottom flas~. The mixture is evaporated to dryness, keeping temperature below 35C, then it is placed into in a thermostatized oven at about 60C until complete dryness. After drying, the product is sieved through a ~6 mesh sieve.
The obtained product is an equimolar mixture (1:1) of verbenone and cyclodextrin (50 g theoretical, 49 g found) .
~XAMPL~ 3 68 g of B-Cyclodextrin are dispersed in 500 ml of distilled water (50 mmoles) and the solution is heated to 65C under stirring, until complete dissolution.
7.51 g of L-verbenone (50 mmoles) are added to this . . ~ . :
, - .
..
W093/06823 2 0 9 7 fi 1 1 PCTIEP.92/02269 solution, with stirring. The mixture is left to cool to room temperature with stirring for 60 minutes, then it is kept at +4C for 6 hours.
The solid phase is recovered by filtration and the solid is placed into a convection oven at 40C for 18 hours, to obtain 64 g of a white powdery product, containing from 11.3 to 12.0% w/w of L-verbenone.
Yield _ 84.65~.
. .
. ~ .
- -, . - .
The present invention relates to the use of a terpene derivative, namely (lS)(-)-4,6,6-trimethylbicyclo-(3,1,1)-hept-3-ene-2-one (hereinafter named verbenone) of formula S O
~ 3 C~
~3 for the preparation of a medicament useful in the treatment of pulmonary emphysema. The invention also relates to a verbenone-cyclodextrin complex and the pharmaceutical compositions containing i~.
Pulmonary emphysema is a progressive respiratory disorder characterized by an abnormal and permanent increase in the volume of the air spaces located distally to the terminal bronchiole, with destruction of the alveolar septum.
A number of experimental and clinical data generally ascribe the emphysema pathogenesis to an unbalance of the protease-antiprotease ratio at the alveolus level. Under normal conditions, alveolar proteases exert an essential activity, which is the lysis of cell fragments, thrombi and particles. Said activity is counterbalanced be endogenous antiproteases ~ 1 antitrypsin [a-lAT], ~-2 macroglob~lin [~-2M], .. ~,,~ . .
.
Bronchial Mucus Inhibitor [B.M.I.]), which yrotect the Dulmonary tissue from the protease action. The loss of the normal balance between proteases and antlproteases can be attributed to three factors: 1) congenital lack in antiproteases (particularly ~-lAT); 2) excessive protease release at the alveolus level; 3) partial inactivation of antiproteases (~-lAT1 in the presence of a normal or enhanced protease production. Said factors are generally worsened by tobacco smoke.
Among alveolar proteases, leukocytal elastase proves to have the most marked digestive activity on all the pulmonary connective components, particularly on elastin, the fibres of which constitute the alveolar septa, thus giving lung the functional elasticity.
Besides the traditional treatments, which cannot remedy the elastin destruction, therapies have been studied to restore elastin synthesis and to protect elastin from elastases action. Satisfactory experimental results were obtained both administering human a-lAT intravenously and stimulating the hepatic synthesis thereof by administration of danatrol, which is a testosterone semi-synthetic derivative. However, both methods suffers from drawbacks, the first method being particularly complex and expensive, whereas the ~econd has antigonadotropinic effects.
Therefore there i5 the need for a medicament exerting an antielastase activity, directly or indi-rectly, and also having poor side-effects.
Now it has been found that verbenone exerts an unpredictable antielestase activity at the lung level, and, dl~e to said activity, it is use~ul in the - ;- .. .. .. .
treatment of emphysema.
Verbenone is a monoterpene derivative havins antiinflammatory and mucolytic activities and it is used as a mucolytic and fluidifying agent in the acute and chronic disorders of the respiratory tract.
As a consequence of further researches, verbenone showed a surprising inhibiting activity on the elastase action, particularly on the neutrophylic one. Said activity was unforeseeable from the pharmacological and clinical studies which evidenced the mucolytic and antiinflammatory activities of verbenone and the use thereof in therapy.
The use of verbenone in the treatment of pulmonary emphysema is extremely advantageous. In fact, pulmonary emphysema can be favoured, or worsened, by the simultaneous presence of other pulmonary disorders, such as bronchitis and/or pneumonia, other infective forms, asthma, which involve obstructions of the respiratory tract; therefore a medicament which can at the same time have antiinflammatory and mucolytic activities and protect elastin of the alveolar septa is obviously useful.
Tests were carried out in vitro to evaluate verbenone activity on the release of neutrophilic elastin from polymorphonuclear granules. Said tests evidenced the marked antielastase activity of verbenone, which activity is not dose-related nor cyto~oxic.
The results from said tests are reported hereinbelow.
Separation of neutro~hilic polymorphonuclears Polymorphonuclear phagocytes were separated starting from heparinized peripheral blood (60 cc) from healthy volunteers or from buffy coats (leukocytes and platelets enrichments).
Neutrophilic polymorphonuclears were separated by means of a 6~ colloidal dextran gradient, then the erythrocytes were lysated with an ammonium chloride solution.
Neutro hilic elastase release p 100 ,ul of cells at a concentration of lxlO7/ml, pretreated with cytochalasin B (5 ~g/ml) for 10', were incubated or 20' at 37C in a microtiter plate in the presence or in the absence of 50 yl of a challenge consisting ~f fMLP (formyl-methionyl-leuc~l-peptide) at a 10-6 molar inal concentration. After said incubation, the plate was centrifuged and 100 ~1 of supernatant were collected and incubated for 60-120' at 37C with a specific substrate for the neutrophilic elastase (methoxysuccinyl-ala-ala-pro-val-p-Na) to a 1 mM final concentration. The reaction was stopped by adding 50 ~1 of 1 N acetic acid and reading was carried out by means of a multiscan photometer at a 405 nm wave length. The values of the absorbance of samples challenged with f.~LP were compared to those from unchallenged cells, to obtain a ~ value corresponding to the amount of neutrophilic elastase released following to challenge. Any interferences of verbenone on the neutrophilic elastase release were tested by addition of different concentrations of the medicament directly to the cell suspension during the test.
,.. ,. . ; . . . . : .
' ' ' ........ ' ' ' ~ ............. . '' ,:
: : -. ,. . .
: . ~ ' . : : ' ' The results are shown in Table 1.
Modulation of the elastase release fro~ neutrophilic polymorphonuclears by addition of verbenone to the cells during the test, at different concentrations.
-% inhibitioncompared to the control Control Verb. 1 mM 90%
Verb. 0,1 mM 86%
Verb. 10 uM 94%
Verb. 1 pM 94%
Verb. 0,1 yM 92%
Verb. 10 nM 89%
Verb. 1 nM 83%
Verb. 0,1 nM 96%
Verb. 10 pM 94%
This marked inhibiting activity is unlikely to be ascribed to a toxic action of the medicament on the cells. In fact the cell viability, as repeatedly evaluated by the tripan bleu exclusion test, turned out to be always higher than 90% after incubation of the cells with verbenone in concentrations varying from 1 mM to lQ pM.
From the industrial point of view, verbenone is used as the active ingredient for the preparation of a medicament useful for the treatment of pulmonary emphysema.
::; . . .. ';` ' ' : .
For the preparation medicaments, verbenone has the drawbacks to be insoluble in water and to have a very unpleasant taste, therefore it cannot be used for aqueous liquid pharmaceutical forms, such as syrups, S oral solutions, nasal drops, aqueous solutions for aerosol and nebulizat on.
It has been found that the verbenone-cyclodextrin complex has high water solubility and, moreover, verbenone unpleasant taste is no more perceptible.
Therefore, the verbenone-cyclodextrin complex is a further object of invention. A further advantage of the complex of the invention is provided by its increased bioavailability.
The pharmaceutical forms comprising the verbenone-cyclodextrin complex can be prepared, for example,according to the methods described in "Remington's Pharmaceutical Sciences Handboo~", Hack Pub. Co., U~A.
Examples of pharmaceutical forms are capsules, pills, tablets, sugar-coated pills, granulates, syrups, drinkable solutions, nasal drops, solutions for aerosol or inhalation, suppositories, injectable solutions or sus~ension~ both for the intravenous and the intramuscular routes.
The dosages will depend on the severity of the disorder, the age, weight and sex of the patient and on the clinician's advise. Examples of dosages comprise single or repeated administrations of the medicament containing verbenone or the verbenone-cyclodextrin complex so as to attain a daily dosage from 10 to 500 mg of the active ingredient. The preparation of the verbenone-cyclod_xtrin complex is illustrated in the .... . .. . . ......... . . . .
~.
:' ' : . . .
: ~ .
W093/06823 2 0 9 7 ~11 PCT/EP92/02269 followin~ examples.
A ~-ciclodextrin amount equal to 0.1 mole is weighed and placed into a mortar of suitable size.
Verbenone ~0.1 mole), previously diluted with 15 ml of 96% ethanol, is added, and the mixture is mixed with a pestle to obtain an homogeneous mass. 20 ml of distilled water are added, and a mixture is obtained which hardens upon mixing. Said mixture is worked in a ~ortar for about 15 minutes, then it is dried in oven thermostatized at 60C.
The obtained complex is in form of a flowing powder which is completely odourless.
Verbenone titre ~ 80% on theoretical.
XA~PL~ 2 5.80 g o verbenone were dissolved in 100 ml of 95C ethanol (to obtain 50 g of a mixture), then they were added to 44.3 g of B-cyclodextrin in a round bottom flas~. The mixture is evaporated to dryness, keeping temperature below 35C, then it is placed into in a thermostatized oven at about 60C until complete dryness. After drying, the product is sieved through a ~6 mesh sieve.
The obtained product is an equimolar mixture (1:1) of verbenone and cyclodextrin (50 g theoretical, 49 g found) .
~XAMPL~ 3 68 g of B-Cyclodextrin are dispersed in 500 ml of distilled water (50 mmoles) and the solution is heated to 65C under stirring, until complete dissolution.
7.51 g of L-verbenone (50 mmoles) are added to this . . ~ . :
, - .
..
W093/06823 2 0 9 7 fi 1 1 PCTIEP.92/02269 solution, with stirring. The mixture is left to cool to room temperature with stirring for 60 minutes, then it is kept at +4C for 6 hours.
The solid phase is recovered by filtration and the solid is placed into a convection oven at 40C for 18 hours, to obtain 64 g of a white powdery product, containing from 11.3 to 12.0% w/w of L-verbenone.
Yield _ 84.65~.
. .
. ~ .
- -, . - .
Claims (5)
1. The use of [(15)(-)-4,6,6-trimethylbicyclo-(3,1,1)-hept-3-ene-2-one] of formula for the preparation of a medicament having antielastase activity.
2. The use of [(lS)(-)-4,6,6-trimethylbicyclo-(3,1,1)-hept-3-en-2-one] for the preparation of a medicament useful for the treatment of pulmonary emphysema.
3. The [(lS)(-4,6,6-trimethylbicyclo-(3,1,1)-hept-3-en-2-one]-cyclodextrin complex.
4. Pharmaceutical compositions containing the complex of claim 3.
5. Compositions according to claim 4, in form of capsules, pills, tablets, sugar-coated pills, granulates, syrups, drinkable solutions, nasal drops, solutions for aerosol or inhalation, suppositories, injectable solutions or suspensions for both the intravenous and intramuscular routes.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
ITMI91A002652 | 1991-10-04 | ||
ITMI912652A IT1251615B (en) | 1991-10-04 | 1991-10-04 | ANTIELASTASIC ACTIVITY MEDICATION. |
Publications (1)
Publication Number | Publication Date |
---|---|
CA2097611A1 true CA2097611A1 (en) | 1993-04-04 |
Family
ID=11360822
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA002097611A Abandoned CA2097611A1 (en) | 1991-10-04 | 1992-09-30 | Verbenone, having antielastase actibity, against pulmonary emphysema |
Country Status (6)
Country | Link |
---|---|
EP (1) | EP0565658A1 (en) |
JP (1) | JPH06506500A (en) |
AU (1) | AU2664492A (en) |
CA (1) | CA2097611A1 (en) |
IT (1) | IT1251615B (en) |
WO (1) | WO1993006823A1 (en) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
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US7157446B2 (en) * | 2003-05-02 | 2007-01-02 | Bristol Myers Squibb Company | Complex of ras-farnesyltransferase inhibitor, a cyclodextrin, and ethanol |
NZ718185A (en) | 2012-06-05 | 2017-12-22 | Univ Korea Res & Bus Found | Pharmaceutical composition containing verbenone derivative for treating or preventing neurodegenerative disease |
CN113603802B (en) * | 2021-08-24 | 2022-10-14 | 华侨大学 | Preparation method and application of verbena polysaccharide |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
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CH625197A5 (en) * | 1976-06-03 | 1981-09-15 | Buskine Sa | Process for preparing verbenone, myrtenal and pinocarveol |
EP0491812A4 (en) * | 1989-09-14 | 1992-11-04 | Australian Commercial Research & Development Limited | Drug delivery compositions |
-
1991
- 1991-10-04 IT ITMI912652A patent/IT1251615B/en active IP Right Grant
-
1992
- 1992-09-30 AU AU26644/92A patent/AU2664492A/en not_active Abandoned
- 1992-09-30 CA CA002097611A patent/CA2097611A1/en not_active Abandoned
- 1992-09-30 EP EP92920465A patent/EP0565658A1/en not_active Withdrawn
- 1992-09-30 WO PCT/EP1992/002269 patent/WO1993006823A1/en not_active Application Discontinuation
- 1992-09-30 JP JP5506603A patent/JPH06506500A/en active Pending
Also Published As
Publication number | Publication date |
---|---|
EP0565658A1 (en) | 1993-10-20 |
JPH06506500A (en) | 1994-07-21 |
IT1251615B (en) | 1995-05-17 |
ITMI912652A1 (en) | 1993-04-04 |
AU2664492A (en) | 1993-05-03 |
WO1993006823A1 (en) | 1993-04-15 |
ITMI912652A0 (en) | 1991-10-04 |
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