CN1222159A

CN1222159A - 6,7-oxygenated steroids and uses related thereto

Info

Publication number: CN1222159A
Application number: CN97195531A
Authority: CN
Inventors: D·L·伯果因; Y·沈; J·M·朗兰斯; C·罗格斯; J·H·L·朝; E·皮尔斯; H·萨拉瑞
Original assignee: University of Alberta; University of British Columbia; Inflazyme Pharmaceuticals Ltd
Current assignee: University of Alberta; University of British Columbia; Inflazyme Pharmaceuticals Ltd
Priority date: 1996-07-11
Filing date: 1997-07-11
Publication date: 1999-07-07
Anticipated expiration: 2017-07-11
Also published as: CN100368425C

Abstract

Steroid compounds having various oxygen substitution on the steroid nucleus are disclosed. A specific functionality present on many of the steroid compounds is oxygen substitution at both of positions 6 and 7. Thus, certain steroids have oxygen substitution at C6 and C7, and some have specific stereochemistries such as 6alpha and 7beta oxygen substitution, and an alpha hydrogen at the 5 position in addition to having 6alpha and 7beta oxygen substitution. Steroids having 3,4-epoxy functionality are also disclosed. In addition, steroids having C17 pyran and delta-lactone functionality, with oxygen substitution at C6 and C7, or at C15, of the steroid nucleus, are disclosed.

Description

6, steroide of 7-oxidation and uses thereof

Related application

The application requires the right of priority of U.S. Provisional Application of submitting on July 11st, 1,996 60/023450 and the U.S. Patent application of submitting on July 12nd, 1,996 08/679642, and above-mentioned two applications are incorporated herein by reference in full.

Technical field

The present invention relates to steroide, particularly 6, steroide of 7-oxidation and uses thereof.

Background of invention

Asthma and allergy have confidential relation, and clinical study has ample evidence to show between the severity of asthma and atopy (allergy) degree strong correlation is arranged.It is believed that the allergy to anaphylactogen is children and the of paramount importance Hazard Factor of adult's asthma, about 90% asthma cases shows allergy.

The increase that is characterised in that SERUM IgE (antibody) level hypersensitive.In being called the process of sensitization, need repeatedly contact allergy former usually to cause that the IgE that abundant B cell produces given antigen or a series of antigen-specifiies excites allergy and asthma subsequently or anaphylaxis.The B cell just can produce the antibody with the mastocyte surface bonding with after antigen contacts.Antigen causes a series of reaction to two antibody crosslinked, causes degranulation and discharges a large amount of media of regulating inflammatory reactions.The medium that discharges in asthma and anaphylaxis or produce comprises histamine, leukotriene, prostaglandin(PG), cytokine and tryptase.

Asthma is characterised in that the overreaction of airway, the stage of attack of bronchospasm and the chronic inflammatory diseases of lung.The obstruction of airway can be in time or the treatment of medicine and reversing.The patient who shows normal airflow may be excessive to multiple natural stimulation such as freezing air, motion, chemical substance and allergenic response.The most common situation that causes asthma reaction is the type to common anaphylactogen, and described anaphylactogen comprises ragweed pollen, showy flowers of herbaceous plants powder, various fungi, dirt mite, cockroach and domestic animal.The symptom of this disease comprises that chest is felt nervous, stridulated, short of breath and cough.The incidence of the mild forms of this disease in American population is up to 10%, and be then higher at the sickness rate of Britain, Australia and New Zealand's report.Although more advanced methods of treatment has been arranged, the incidence of asthma and mortality ratio are all increasing in the world, have turned over one times in the past 20 years.

Airway is very complicated to the reaction of anaphylactogen, comprises early stage asthma reaction (EAR), and this is reflected at after the haptic stimutus 20-30 minute and peaks, and it is characterized in that bronchial contraction and eliminates after 1.5-2 hour usually.Late asthmatic response (LAR) takes place after 3-8 hour in initial contact usually, comprises bronchoconstriction and inflammation and oedema occur in lung tissue.This inflammation can become chronic usually, the infiltration to lung of epithelial damage and inflammatory cell such as eosinophil and neutrophil occurs.

Treat the method for asthma at present

Glucocorticosteroid (steroide) is the most effective extended regimen of treatment asthma.Oral steroide is very ineffective for control acute asthma outbreak, and because the employing of induction type steroide, and the chronic application of its control asthma also seldom.Even owing in mild asthma, also exist the inflammation of airway, and therefore, induction type steroide even just having brought into use in early days in pharmacological agent.Though the induction type steroide is very effective, side effects limit its application, thereby often adopt combination therapy.Combination therapy is divided into following several sections: antiphlogiston (for example suck and oral steroide), bronchodilator (β for example ₂-agonist, xanthine derivative, anti-cholinergic thing) and medium inhibitor (for example Cromoglycic Acid compounds and leukotriene antagonist).

Cromoglycic Acid compounds (for example sodium cromoglycate and nedocromil) can and prevent bronchial overreaction in the release of vitro inhibition histamine, and side effect seldom.They are oral invalid and do not have a bronchiectatic activity.Although, need chronic treatment (a couple of days) to reach best antiphlogistic effects usually as long as the Cromoglycic Acid compounds just can produce beneficial effect to kinetic asthma when motion administration in preceding 10 minutes.The Cromoglycic Acid compounds at most only can be calculated to effectively for moderate to serious asthma.

Glucocorticosteroid (steroide) is very effective to pneumonia, is to be used for the treatment of asthma and the most effectively medicine hypersensitive at present.They can suppress arachidonic acid metabolite (leukotriene and prostaglandin(PG)) and production of cytokines in mastocyte.According to the severity of disease, can in 4 hours, occur the reaction of induction type steroide or general steroide, but also may need a couple of days.When not carrying out normal chronic treatment, symptom can recur usually.Use the side reaction of induction type steroide to comprise dysphonia, local excitation and oral candidiasis (a kind of fungi infestation) continuously.More the induction type steroide of high dosage can cause the inhibition of HPA-axle, and described HPA-axle is responsible for regulating serum cortisol level, metabolism, stress reaction, CNS function and immunity.Use the induction type steroide or the oral steroide of high dosage can cause more serious side reaction continuously: the severe inhibition of hpa axis, cause immune influence, hypertension, osteoporosis, peptide ulceration, growthretardation, behavior disorder, dysgenesia, cataract and hemopathy.

Beta-2-agonists can reverse the bronchospasm that produces in the asthma attack process, the outbreak of reacting is had medium activity.The route of administration of this medicine can be different with acting duration.Prolong and use these reagent tolerance can occur, thereby cause weakening this treatment reaction itself.These compounds are for not effect of inflammatory reaction itself.

Xanthine derivative is the inhibitor of cyclic amp phosphodiesterase diesterase, also is used for the bronchiectasis therapy.Though xanthine is effective, its activity is subjected to multiple factor to comprise the influence of food, age, smoking etc.Therapeutic domain relative narrower, its side reaction comprise gastrointestinal tract disease, CNS disorder, headache, anxiety and irregular pulse.The importance of treatment inflammation makes in treatment fewer and feweri to xanthic application in asthma and allergy.

Anti-cholinergic thing such as ipratropium bromide are used to block because the contraction of the caused bronchial smooth muscle of release neurotransmitters vagusstoff.These medicines are the most effective for the treatment chronic obstructive pulmonary disease, only reported some positive effect in asthma.A large amount of side reactions of these medicines comprise uroschesis, dry, tachycardia, feel sick, vomiting, flushing and hypertension.

The 5-lipoxidase inhibitor suppresses the generation of leukotriene, and leukotriene antagonist can stop leukotriene to be had an effect, and described leukotriene is the medium of the powerful bronchospasm that discharges in the asthma reaction process.Use the leukotriene synthetic inhibitor this means the function that in some patient group, to pay close attention to liver with the increase of liver enzyme.Leukotriene inhibitors shows the activity suitable with the Cromoglycic Acid compounds, and its activity is equivalent to the cortin of low dosage.

Generally speaking, moderate is considerably less to a serious asthmatic patient applicable medicine.The safe drugs effect a little less than, and effectively medicine has disadvantageous side reaction and need extensively monitor the patient.Therefore, need the safety and the therapeutical agent of relieving asthma and allergic symptom effectively.The invention provides these beneficial effects described in the literary composition.

Summary of the invention

On the one hand, the invention provides the compound of following formula:

Comprise its pharmacy acceptable salt and solvate, wherein:

C1, C2, C3, C4, C11, C12, C15 and C16 independently of one another by (a) and (b) both one of replace:

(a) one of following formula :=O ,=C (R ⁴) (R ⁴) ,-C (R ⁴) (R ⁴) (C (R ⁴) (R ⁴)) _n-and-(O (C (R ⁴) (R ⁴)) _nO)-, wherein, n is 1 to about 6;

(b) two in the following formula, each is independently selected from :-X ,-R ⁴With-OR ¹

C5, C6, C7, C8, C9, C10, C13 and C14 independently of one another by-X ,-R ⁴Or-OR ¹One of replace;

C17 is by (c), (d), (e), (f), (g), (h) with any replacement (i):

(c)=C (R ²) (R ³), but C14 during by methyl substituted except;

(d)-R ⁵With-OR ⁶, as long as work as C10 by methyl substituted, C5 directly is not connected with oxygen, wherein R ⁵And R ⁶Can form direct bond together, thereby C17 is carbonyl, perhaps R ⁵And R ⁶Form 3-6 unit's cyclic ethers or 4-6 membered ring lactone with C17; Otherwise, R ⁵Be R ⁴Or-OR ⁶, R ⁶Be R ¹Or R ⁴

(e) one of following formula :=O ,=C (R ⁴) (R ⁴) ,-C (R ⁴) (R ⁴) (C (R ⁴) (R ⁴)) _n-and-(O (C (R ⁴) (R ⁴)) _nO)-, wherein, n is 1 to about 6, as long as satisfy one of following condition:

ⅰ) C5 is replaced by hydrogen with the α configuration, and C3 is not connected with oxygen;

If ⅱ) C10 is by methyl substituted, the A ring is aromatic ring, and then C13 and C14 be not all by methyl substituted;

If ⅲ) C3 is connected with Sauerstoffatom with C4, and C6-OR ¹Substituting group has the α configuration, and C7-OR ¹Substituting group has beta comfiguration, and then C17 is not by any replacement in the following radicals:

ⅳ) C3 and C4 all are connected with identical Sauerstoffatom, thereby form an oxyethane ring, and condition is: when C5 have hydroxyl or-OR ¹During substituting group, C7 does not have the carbonyl substituted base;

(f) two kinds in the following substituting group, it can be independently selected from :-X ,-R ⁴With-OR ¹, as long as above-mentioned condition ⅰ), ⅱ), ⅲ) or one of ⅳ) can satisfy;

(g) ring structure of following formula

Wherein, G be-C (=O)-,-CH (OR ¹)-,-C (R ⁴) (OR ¹)-or-C (OR ¹) (OR ¹)-, be not as long as C3 and C4 are replaced by the hydroxyl of hydroxyl or protection simultaneously;

(h) two hydrogen atoms are not as long as C3 is by carbonyl substituted;

(ⅰ) hydrogen atom and one are selected from C ₁-C ₃₀Alkyl and C ₁-C ₃₀The group of halo alkyl does not comprise-CH (CH ₃) (CH ₂) ₃CH (CH ₃) ₂

A, B, C and D ring can be complete saturated rings, fractional saturation ring or complete unsaturated ring independently;

R ¹Be H, thereby or a kind of blocking group make-OR ¹Be the oh group of protection, wherein, ortho position-OR ¹Group can form the ring structure of protection vicinal hydroxyl groups together, and geminal-OR ¹Group can form the ring structure of protection carbonyl together, and condition is one-OR on C6 and C7 position ¹Or two-OR ¹Represent the carbonyl of carbonyl or protection;

Each R ², R ³And R ⁴All be independently selected from H and C ₁-C ₃₀Organic residue, this organic residue can optionally comprise at least one heteroatoms, and described heteroatoms is selected from boron, halogen, nitrogen, oxygen, silicon and sulphur, wherein, two geminal R ⁴Group can form a ring with the carbon atom that is connected with them; With

X represents fluorine, chlorine, bromine and iodine.

In preferred embodiments, compound is a following formula: compound

Comprise its pharmacy acceptable salt and solvate, wherein:

C1, C2, C3, C4, C11, C12, C15 and C16 are replaced by following radicals independently of one another:

(a) one of following formula :=O ,=C (R ⁴) (R ⁴) ,-C (R ⁴) (R ⁴) (C (R ⁴) (R ⁴)) _n-and-(O (C (R ⁴) (R ⁴)) _nO)-, wherein, n is 1 to about 6; Or

C5, C8, C9, C10 and C13 independently of one another by-X ,-R ⁴Or-OR ¹One of replace;

C14 is-X ,-OR ¹Or except that methyl-R ⁴

A, B, C and D ring can saturated fully independently, fractional saturations or unsaturated fully;

X represents fluorine, chlorine, bromine and iodine.

In another preferred embodiment, compound is a following formula: compound

Comprise its pharmacy acceptable salt and solvate, wherein:

(b) two in the following formula: each is independently selected from :-X ,-R ⁴With-OR ¹

C5, C8, C9, C10, C13 and C14 independently of one another by-X ,-R ⁴Or-OR ¹One of replace;

Each R ⁴All be independently selected from H and C ₁-C ₃₀Organic residue, this organic residue can optionally comprise at least one heteroatoms, and described heteroatoms is selected from boron, halogen, nitrogen, oxygen, silicon and sulphur, wherein, two geminal R ⁴Group can form a ring with the carbon atom that is connected with them; With

R ⁵And R ⁶Can form direct bond together, thereby C17 is carbonyl, perhaps R ⁵And R ⁶Form 3-6 unit's cyclic ethers or 4-6 membered ring lactone with C17; Otherwise, R ⁵Be R ⁴Or-OR ⁶, R ⁶Be R ¹Or R ⁴With

X represents fluorine, chlorine, bromine and iodine.

Condition is, as C10 during by methyl substituted, then C5 directly is not connected with Sauerstoffatom.

In another preferred embodiment, compound is a following formula: compound

Comprise its pharmacy acceptable salt and solvate, wherein:

C1, C2, C4, C11, C12, C15, C16 and C17 are replaced by following radicals independently of one another:

C8, C9, C10, C13 and C14 independently of one another by-X ,-R ⁴Or-OR ¹One of replace;

C3 quilt=C (R ⁴) (R ⁴) and-C (R ⁴) (R ⁴) (C (R ⁴) (R ⁴)) _n-one of replace, wherein n is 1 to about 6, perhaps by-X and-R ⁴In two replacements, condition is that C3 is not connected with Sauerstoffatom;

R ¹Be H, thereby or a kind of blocking group make-OR ¹Be the oh group of protection, wherein, ortho position-OR ¹Group can form the ring structure of protection vicinal hydroxyl groups together, and contraction position-OR ¹Group can form the ring structure of protection carbonyl together, and condition is one-OR on C6 and C7 position ¹Or two-OR ¹Represent the carbonyl of carbonyl or protection;

X represents fluorine, chlorine, bromine and iodine.

In another preferred embodiment, compound is a following formula: compound

Comprise its pharmacy acceptable salt and solvate, wherein:

C1, C2, C3, C4, C11, C12, C15, C16 and C17 are replaced by following radicals independently of one another:

Condition is: (a) C10 and C13 be simultaneously by methyl substituted, and (b) as C10 during by methyl substituted, then C14 is not by methyl substituted;

A, B, C and D ring can be complete saturated rings, fractional saturation ring or complete unsaturated ring independently, and condition is that the A ring is non-aromatic ring;

R ¹For thereby H or a kind of blocking group make-OR ¹Be the oh group of protection, wherein, ortho position-OR ¹Group can form the ring structure of protection vicinal hydroxyl groups together, and geminal-OR ¹Group can form the ring structure of protection carbonyl together, and condition is one-OR on C6 and C7 position ¹Or two-OR ¹Represent the carbonyl of carbonyl or protection;

X represents fluorine, chlorine, bromine and iodine.

In another preferred embodiment, compound is a following formula: compound

Comprise its pharmacy acceptable salt and solvate, wherein:

C1, C2, C11, C12, C15, C16 and C17 are replaced by following radicals independently of one another:

Condition is that C17 is not by any replacement in the following radicals:

C5, C9, C10, C13 and C14 independently of one another by-X ,-R ⁴Or-OR ¹One of replace;

C8 quilt-X or-R ⁴Replace, preferably directly be not connected with oxygen;

R ¹Be H, thereby or a kind of blocking group make-OR ¹Be the oh group of protection, wherein, ortho position-OR ¹Group can form the ring structure of protection vicinal hydroxyl groups together, and geminal-OR ¹Group can form the ring structure of protection carbonyl together;

X represents fluorine, chlorine, bromine and iodine.

In another preferred embodiment, compound is a following formula: compound

Comprise its pharmacy acceptable salt and solvate, wherein:

Condition is that C3 and C4 are not replaced by the hydroxyl of hydroxyl or protection simultaneously, are not preferably replaced by Sauerstoffatom simultaneously;

G is-C (=O)-,-CH (OR ¹)-,-C (R ⁴) (OR ¹)-or-C (OR ¹) (OR ¹)-;

X represents fluorine, chlorine, bromine and iodine.

In another preferred embodiment, compound is a following formula: compound

Comprise its pharmacy acceptable salt and solvate, wherein:

X represents fluorine, chlorine, bromine and iodine;

Condition is, when C5 have hydroxyl or-OR ¹During substituting group, C7 does not have the carbonyl substituted base.

In another preferred embodiment, compound has and is selected from following structural formula:

Comprise its pharmacy acceptable salt and solvate, wherein:

C1, C2, C3, C4, C11, C12 and C16 independently of one another by (a) and (b) both one of replace:

C5 is replaced by hydrogen atom;

C6, C7, C8, C9, C10, C13 and C14 independently of one another by-X ,-R ⁴Or-OR ¹One of replace; With

C17 quilt (c), (d), (e) or (f) replacement:

(c) two are selected from following substituting group: hydrogen, halogen, remove-CH (CH ₃) (CH ₂) ₃CH (CH ₃) ₂Outside C ₁-C ₃₀Saturated hydrocarbyl, halo C ₁-C ₃₀Saturated hydrocarbyl, C ₁-C ₃₀Unsaturated alkyl, halo C ₁-C ₃₀Unsaturated alkyl;

(d) be selected from=C (R ⁴) (R ⁴) substituting group, condition is that C14 is not by methyl substituted;

(e) at least a substituting group that contains Sauerstoffatom, it is selected from=O ,-(O (C (R ⁴) (R ⁴)) _nO)-,-OH and-OR ¹, wherein, n is 1 to about 6;

(f) substituting group of at least a nitrogen atom, it is selected from-N (R ⁴) (R ⁴), wherein, two R ⁴Group can form one or more rings with nitrogen-atoms, thereby the substituting group of nitrogen atom comprises the heterocyclic group of nitrogen atom; Wherein

R ¹Be H, or a kind of blocking group, thereby make-OR ¹Be the oh group of protection, wherein, be attached on the adjacent carbons-OR ¹Group can form the ring structure of two hydroxyls of protection together;

Each R ⁴All be independently selected from H and R ⁵

R ⁵Be C ₁-C ₃₀Organic residue, this organic residue can optionally comprise at least one heteroatoms, and described heteroatoms is selected from boron, halogen, nitrogen, oxygen, silicon and sulphur, wherein, two geminal R ⁵Group can form a ring with the carbon atom that is connected with them; With

X represents fluorine, chlorine, bromine and iodine.

On the other hand, the invention provides a kind of pharmaceutical composition, it comprises above-mentioned any compound and pharmaceutically acceptable carrier or thinner.

On the other hand, the invention provides a kind of pharmaceutical composition, it comprises a kind of compound and pharmaceutically acceptable carrier or thinner, and this compound is a following formula: compound

Comprise its pharmacy acceptable salt and solvate, wherein:

C1, C2, C3, C4, C11, C12, C16 and C17 are selected from (a) independently of one another or substituting group (b) replaces, wherein

(a) represent one of following formula :=O ,=C (R ⁴) (R ⁴) ,-C (R ⁴) (R ⁴) (C (R ⁴) (R ⁴)) _n-and-(O (C (R ⁴) (R ⁴)) _nO)-, wherein, n is 1 to about 6;

(b) representative-X ,-R ⁴With-OR ¹In two, each is all selected independently;

R ¹Be H, thereby or a kind of blocking group make-OR ¹Be the oh group of protection, wherein, C6 and C7-OR ¹Group can form the ring structure of two hydroxyls of protection together;

Each R ⁴All be independently selected from H and R ⁵

R ⁵Be C ₁-C ₃₀Organic residue, this organic residue can optionally comprise at least one heteroatoms, and described heteroatoms is selected from boron, halogen, nitrogen, oxygen, silicon and sulphur, wherein, two geminal R ⁴Group can form a ring with the carbon atom that is connected with them; With

X represents fluorine, chlorine, bromine and iodine;

Condition is that C15 does not link to each other with Sauerstoffatom.

On the other hand, the invention provides above-claimed cpd (any or its mixture) and be used for the treatment of asthma, allergy in production, comprise the inflammation and/or the thrombosis of sacroiliitis, or be used for the treatment of and NF _KPurposes in the medicine of the illness that the rising of B level is relevant.

On the other hand, the invention provides a kind of method for the treatment of asthma, comprise the aforesaid compound or its salt or the pharmaceutical composition that need the patient of this treatment significant quantity.

On the other hand, the invention provides a kind of treatment method hypersensitive, comprise the aforesaid compound or its salt or the pharmaceutical composition that need the patient of this treatment significant quantity.

On the other hand, the invention provides a kind of treatment because the method for the inflammation that sacroiliitis causes comprises the aforesaid compound or its salt or the pharmaceutical composition that need the patient of this treatment significant quantity.

On the other hand, the invention provides the thrombotic method of a kind of treatment, comprise the aforesaid compound or its salt or the pharmaceutical composition that need the patient of this treatment significant quantity.

On the other hand, the invention provides a kind of treatment and NF _KThe method of the illness that the rising of B level is relevant comprises the aforesaid compound or its salt or the pharmaceutical composition that need the patient of this treatment significant quantity.

On the other hand, the invention provides a kind ofly, introduce the method for the outer olefin group of ring in the 7-titanium dioxide steroide, comprise the compound that a kind of formula (10) are provided to 6, the Witting reagent of formula (10) compound and formula (11) is reacted in the presence of alkali, obtain the olefin(e) compound of a kind of formula (12)

Wherein, formula (10) and formula (12) compound all comprise its pharmacologically acceptable salt and its solvate, wherein:

C1, C2, C3, C4, C11, C12, C15 and C16 are independently of one another by (a) with one of (b) replace:

R ¹Be H, or a kind of blocking group, thereby make-OR ¹Be the oh group of protection, wherein, ortho position-OR ¹Group can form the ring structure of protection vicinal hydroxyl groups together, and geminal-OR ¹Group can form the ring structure of protection carbonyl together, and condition is one-OR on C6 and C7 position ¹Or two-OR ¹Represent the carbonyl of carbonyl or protection;

Each Ra, Rb and R ⁴All be independently selected from H and C ₁-C ₃₀Organic residue, this organic residue can optionally comprise at least one heteroatoms, and described heteroatoms is selected from boron, halogen, nitrogen, oxygen, silicon and sulphur, wherein, two geminal R ⁴Group can form a ring with the carbon atom that is connected with them; With

X represents fluorine, chlorine, bromine and iodine; It can be selected under each situation independently.

On the other hand, the invention provides a kind of with 6 α, 7 β-titanium dioxide group is introduced the method in the steroide, comprise that providing a kind of has carbonyl and at the steroide that C5 and C6 interdigit have the formula (13) of two keys, be hydroxyl with carbonyl reduction, again with two key hydroborations in the C7 position, obtain hydroxyl in the C6 position, thereby the hydroxyl of C6 position has α-configuration, and the hydroxyl of C7 position has beta configuration

Wherein, the compound of formula (13) and formula (14) comprises its pharmacy acceptable salt and its solvate, wherein:

C1, C2, C3, C4, C11, C12, C15, C16 and C17 are independently of one another by (a) with one of (b) replace:

X represents fluorine, chlorine, bromine and iodine.

On the other hand, the invention provides a kind of method that is used for introducing hydroxyl in the C3 position of steroid nucleus stereoselectivity, this method comprises provides a kind of compound that has the formula (15) of carbonyl on the C3 position, with reductive agent carbonyl reduction is become hydroxyl, thus at least a compound of the formula of obtaining (16) and (17)

Wherein, the compound of formula (15), (16) and (17) respectively comprises its pharmacy acceptable salt and its solvate, wherein:

C1, C2, C4, C11, C12, C15, C16 and C17 are independently of one another by (a) with one of (b) replace:

X represents fluorine, chlorine, bromine and iodine.Detailed Description Of The Invention

The present invention relates to various steroid derivatives with specific function base of describing in detail in the text.Compound exhibits as herein described goes out to suppress the mastocyte degranulation, suppress the bronchospasm (acute phase) that anaphylactogen causes and suppress the effect of the pneumonia (late period) that anaphylactogen causes, so can be used as the good control agent of asthma and anaphylaxis.These compounds represented a kind of material of new series, this material has potential treatment benefit for treatment asthma and allergy, has efficient, broad spectrum of activity and low side effect.

For ease of the new feature of identification The compounds of this invention, shown in the following structural formula 1 of the unsubstituted steroid nucleus of single-minded its each ring carbon atom of Digital ID.This numbering system will be used in this article all the time.

Structural formula 1

Compound of the present invention comprises two unsymmetrical carbons at least, thereby has enantiomorph and diastereomer isomer.Except as otherwise noted, the present invention includes the following formula compound of all enantiomorphs and diastereomer form.The mixture of the different compounds of the mixture of pure stereoisomers, enantiomorph and/or diastereomer and following formula includes within the scope of the invention.

Synthetic method described herein particularly when combining with the common sense of this area, provides useful guidance to those skilled in the art, with synthetic, separation and purifying preferred compound of the present invention and other analogue compounds.If necessary, can as forming diastereomer, carry out recrystallization again, from multi-form mixture, obtain indivedual enantiomorphs according to known method for splitting.

The compound of following formula can be solvate forms or its pharmacy acceptable salt form, as acid salt.This salt comprise hydrochloride, vitriol, phosphoric acid salt, Citrate trianion, fumarate, mesylate, acetate, tartrate, maleate, lactic acid salt, mandelate, salicylate, succinate and other salt well known in the art.

Compound of the present invention can be by making up with the preparation composition with pharmaceutically acceptable carrier or thinner.Suitable carrier or thinner comprise physiological saline.Those skilled in the art will appreciate that steroide or one or more steroides and one or more non-steroids that composition of the present invention can comprise more than one make up.

Being present in the many kinds of specificity officials on the steroide of the present invention can be all to have the oxygen substituting group at 6 and 7.Therefore, some steroide of the present invention has the oxygen replacement mode shown in structural formula 2.Another feature of some this steroid-like is that it has specific stereochemistry.For example, shown in structural formula 3, have steroide that 6 α and 7 β oxygen replace and shown in structural formula 4, remove and have the replacement of 6 α and 7 β oxygen, the steroide that also has α hydrogen on 5 all falls within protection scope of the present invention.

Structural formula 2

Structural formula 3

Structural formula 4

In structural formula 2,3 and 4, each Sauerstoffatom that is connected with carbon 7 with carbon 6 simultaneously and " R ¹" group links to each other.R ¹Group is hydrogen or hydroxyl protecting group.Suitable protecting group is stated in following document: Greene, " protecting group in the organic chemistry ", John wiley ﹠amp; Sons, NewYork NY (1981).When the compound of structural formula 2-4 comprises ortho position-OR ¹Group (that is two-OR on adjacent carbons, ¹Group) time, ortho position-OR ¹Group can form the ring structure of protection vicinal hydroxyl groups together.Ketal is the ortho position-OR of protection ¹Examples of groups.Geminal-OR ¹Group (that is two-OR on same carbon atom, ¹Group) can form the ring structure of protecting carbonyl together.Ketal is an example of this ring structure.Should be appreciated that one-OR on C6 and C7 position ¹Or two-OR ¹Represent the carbonyl of carbonyl or protection, therefore, at C6 and C7 position, R ¹Can be the direct bond between Sauerstoffatom and the carbon atom that links to each other with Sauerstoffatom (C6 or C7).

Steroide of the present invention can at C8 and/or the C9 position has or for α or for the stereochemical substituting group of β.Hydrogen atom on steroide C8 of the present invention position is generally beta comfiguration.In addition, the preferred steroide of the present invention can have the stereomeric methyl substituents of β at C10 and/or C13 position.When C15 was not ketone, compound of the present invention preferably had the stereomeric C14 hydrogen for α.Preferably have in the substituent steroide in the C17 position in the present invention, the C17 substituting group has β stereoisomerism configuration.

In β-ring, have 6 according to structural formula 2, the steroide of 7-titanium dioxide group can commercially available have α by many in the A-ring, the steroide precursor of beta-unsaturated carbonyl synthesizes, comprise 4-androstene-3,17-diketone (following compound 1) and dehydroepiandrosterone (following compound 247).These special steroide precursors can be from Steraloids Inc., wilton, and N.H. obtains.Other is suitable has C3 oxygen functional group and Δ ⁵The steroide precursor of carbon-to-carbon double bond can (Milwaukee WI) obtains from Aldrich Chemical Co. for example.

From 4-androstene-3, the synthetic order of illustrative that the 17-diketone prepares structural formula 2 compounds is summarized in following reaction scheme 1.

Reaction scheme 1

Beginning is by carbonyl-protection base protection 4-androstene-3, the carbonyl functional group of 17-diketone.Shown in reaction scheme 1, this can be by making compound 1 and (CH ₂OH) ₂With the reaction of the benzole soln of p-TsOH, and then carbonyl changed into ketal group and finish.Other suitable carbonyl-protection base is stated in following document: Greene, " blocking group in the organic chemistry ", John Wi1ey ﹠amp; Sons, New, York, NY (1981).Under the acidic conditions that forms the protection ketone groups, can produce of the following property migration of C4-C5 carbon-to-carbon double bond to the C5-C6 position, finally form compound 2.

C5-C6 carbon-to-carbon double bond to compound 2 carries out the allylic oxidation, carbonylate oxygen on the C7 position, thus form compound 3.Multiple oxygenant and experiment condition can be used for the allylic oxidizing reaction, comprise chromium trioxide/3 title complex, chloro chromic acid pyridine (PCC), dichromic acid pyridine (PDC) or RuCl ₃Add tertbutyl peroxide.

The C7 ketone that forms reduces with suitable reductive agent, obtains the hydroxyl functional base of C7 position, shown in compound 4.In several metal hydride reducing agents any can be used for this purpose, comprises sodium borohydride or LiAlH ₄Usually, by the minimum of the hydride ion attack steroide face that is obstructed, make the reduction of C7 ketone and produce β-OH configuration.Then, the most handy hydroxyl protecting group of C7 oh group such as tertiary butyl dimethylsilane (TBDMS) protection obtain protected vinyl carbinol, are compound 5.Other suitable hydroxyl protecting group is listed in Greene, and ibid for document.

Before or after protection C7 hydroxyl,, by ring-opening reaction, can on C6, introduce oxygen again by such as hydroboration/oxidizing reaction or epoxidation reaction.For example, the Δ of compound ⁵Carbon-to-carbon double bond can comprise metachloroperbenzoic acid, trifluoroperacetic acid or 3 with any, and 5-dinitrobenzene benzoyl hydroperoxide carries out epoxidation at interior peracid, obtains the epoxide as compound 6.Usually, the epoxide of introducing has α-configuration, because be the minimum of the attack steroide ring structure face that is obstructed.The ring-opening reaction of epoxide can be carried out under 60 ℃ and acidic conditions, as 80% acetic acid aqueous solution.Crude mixture inclusion compound 7 (have vinyl carbinol in the C6 position, have α-configuration) and its C7 position silyl derivative.With tetrahydrofuran (THF) (THF) solution-treated of this crude mixture, obtain simplification compound 7 with tetrabutyl ammonium fluoride (TBAF).In addition, adopt suitable borane complexes to Δ ⁵Carbon-to-carbon double bond carries out hydrogenation, and the reagent that re-uses such as alkaline hydrogen peroxide carries out the hydroxyl that oxidation also can be introduced α-configuration in the C6 position.

Compound 7 is for having the illustrative compound of structural formula 2 and 3 oxidised forms.The method that compound 1 is changed into structural formula 2 and/or 3 compounds is suitable for variously having α, the compound of beta-unsaturated carbonyl group at the A of steroide in the ring usually.Other structural formula 2 and/or 3 compound can be by obtaining modifying such as the dihydroxy compound of compound 7.In this case, must protect each C6 and C7 hydroxyl, the method that realizes this protection will be described in the back.

Compound 7 or its analogue can change into the compound of structural formula 4.Say that simply this process can realize like this: protection C6 and C7 hydroxyl and C17 carbonyl, then with Δ ⁴The carbon-to-carbon double bond reduction.Lithium in ammonia/THF is the example of the reductive agent that suits.This reduction process obtains a kind of enolate, and it can adopt suitable electrophilic reagent such as trimethylsilyl chloride or chloro di(2-ethylhexyl)phosphate ethyl ester to catch.

The example of this conversion process is shown in the reaction scheme 2.Therefore, available 2, (1S)-(+) of 3-Propanal dimethyl acetal and catalytic amount-10-camphorsulfonic acid (CSA) is handled the protection of going up hydroxyl with C6 that realizes compound 7 and C7, obtains acetone solvate 8.The C17 carbonyl of compound 8 can be by converting it into hydroxyl protection, and then the protection hydroxyl.Can use sodium borohydride in the methyl alcohol to finish the chemical selective reduction of C17 carbonyl, obtain compound 9, compound 9 and suitable hydroxyl protecting group such as tert-butyldimethylsilyl chloride are reacted, obtain silyl ether compound 10.Compound 10 can react with the lithium among ammonia/THF, with the reaction of chloro di(2-ethylhexyl)phosphate ethyl ester, obtains compound 11 again.Compound 11 has 5 α hydrogen and C6 and C7 dihydroxyl group, is the compound of representational structural formula 4 therefore.

Reaction scheme 2

According to one aspect of the present invention, outside the C17 position has ring alkene is provided and has all had the alkene steroide of Sauerstoffatom in C6 and C7 position.In one embodiment, this alkene steroide has structural formula 5, comprises enantiomorph or geometrical isomer that it is independent, further comprises its solvate or pharmacy acceptable salt.Structural formula 5 is defined as follows:

The compound of following formula

Comprise its pharmacy acceptable salt and solvate, wherein:

C14 independently by-X, except that methyl-R ⁴Or-OR ¹One of replace;

X represents fluorine, chlorine, bromine and iodine.

On C17, provide a kind of process of exocyclic double bond to be easy to finish by Wittig reaction by being raw material with the C17 carbonyl compound.Steroide of the present invention with C17 carbonyl functional group is easy to obtain, and for example, as the compound 7 of reaction scheme 1 preparation, perhaps the synthetic order of being summarized by following reaction scheme 3 prepares (comparing with reaction scheme 2) by compound 10.

Reaction scheme 3

The A ring of compound 10 can be reduced so that the C3 carbonyl becomes unique functional group of A ring.Reaction scheme 3 has illustrated that wherein compound 10 is used in the lithium reduction among liquefied ammonia and ether solvents such as ether or the THF, obtains the mixture of compound 12 and compound 13 in order to realize the two steps order of this reduction process.Then, this mixture is carried out oxidation, obtain simplification compound 13 with suitable oxygenant such as PDC.Use again LS-Selectride  (Aldrich Chemical Co., Milwaukee, WI) or other selective reduction agent with compound 13 reduction, stereochemical compound 14 shown in obtaining having.

Then, use diacetyl oxide and pyridine that 3 Alpha-hydroxies of compound 14 are obtained compound 15 with acetic ester protection.Other suitable hydroxyl protecting group can be used to replace aceticoceptor.Known in the artly be used to remove under the standard conditions of silyl protecting group, for example use tetrabutyl ammonium fluoride (TBAF), remove the silyl protecting group of C17 position, obtain C17 oxy-compound such as compound 16.Under conventional oxidizing condition, as using the DMSO solution and the triethylamine of oxalyl chloride, can be carbonyl with the C17 hydroxyl oxidize, obtain ketone compound 17.

Compound 17 can be used in the multiple olefination, comprises in the reaction of Wittig type, to obtain having in the C17 position compound of the structural formula 5 of alkene.For example, compound 17 can react with the ethyl triphenyl phosphonium bromide, obtains ethylidene compound 18.Other raw ketone can be used for obtaining other has exocyclic double bond in the C17 position steroide.

As previously mentioned, the compound that comprises carbonyl (or those are easy to change into the functional group of carbonyl) in the C17 position can change into the compound that has carbon-to-carbon double bond in the C17 position by the Wittig chemical reaction.For example, as following reaction scheme 4 was listed, in the reaction of step First Five-Year Plan, compound 19 can change corresponding C 17 ethylidene compounds 23 into.2 α of compound 19,3 beta-dihydroxyl functional groups can protect (for example, using 2, the N of 2-Propanal dimethyl acetal and camphorsulfonic acid (CSA), dinethylformamide (DMF) solution) with hydroxyl protecting group, obtain the compound as compound 20.The deprotection of C17 hydroxyl can use the reaction conditions (can be used in the TBAF among the THF herein) that is applicable to concrete hydroxyl protecting group, again the hydroxyl oxidize that forms (as is used in CH ₂Cl ₂In PDC), obtain comprising the compound 21 of C17 ketone.With the toluene solution reaction of compound 21, obtain compound 22 with Witting reagent such as ethyl triphenyl phosphonium bromide and uncle's fourth potassium oxide.Behind the hydroxyl deprotection in alkene 22, obtain tetrahydroxy compound 23.

Reaction scheme 4

In some cases, the step that before the derivatize of C17 position, needs protection.For example, in compound 24 (according to reaction scheme 14 preparations), C3 ketone should at first be protected (referring to reaction scheme 5) before the C17 position changes.At first, carry out acetylization reaction (for example, using the pyridine solution of diacetyl oxide) again, obtain C3, C5-acetoxyl group derivative 25 compound 24 reduction (for example reacting) by ethanolic soln with sodium borohydride.Similar with reaction scheme 4 described processes, carry out the oxidation and the Wittig chemical reaction of C17 position, obtain compound 27.Carry out the deprotection (adopting 80% acetate usually to remove the ketal group of compound 27) of C6 and C7 hydroxyl subsequently, obtain compound 28, it comprises the outer Δ of ring ¹⁷Alkene.

Reaction scheme 5

According to another aspect of the present invention, provide steroide with C17 oxide group and C6 and C7 position oxide group.In one embodiment, this steroide has structural formula 6, comprises enantiomorph or geometrical isomer that it is independent, further comprises its solvate or pharmacy acceptable salt.Structural formula 6 is defined as follows:

Comprise its pharmacy acceptable salt and solvate, wherein:

R ⁴And R ⁶Can form direct bond together, thereby C17 is carbonyl, perhaps R ⁵And R ⁶Form 3-6 unit's cyclic ethers or 4-6 membered ring lactone with C17; Otherwise, R ⁵Be R ⁴Or-OR ⁶, R ⁶Be R ¹Or R ⁴With

X represents fluorine, chlorine, bromine and iodine.

Many examples of the compound of structural formula 6 and its synthetic method provide in front.Compound 7,8,9,10,11,12,13,14,15,16,17,19,20,21,24,25 and 26 is the representative compounds of structural formula 6.Other compound of many structural formulas 6 and synthetic method thereof all provide in conjunction with other compound of the present invention in this article.Thereby those of ordinary skill in the art can disclosure according to the present invention prepares the compound of many kinds of structural formulas 6.

The compound of structural formula 6 can have oxygen and/or hydrocarbyl substituent in the C1 position.Below providing the C1 position at structural formula 6 compounds that is shown in reaction scheme 6,7 and 8 provides the illustrative synthetic method of oxygen and/or hydrocarbyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 compound of C1 position aerobic and/or hydrocarbyl substituent provides the oxygen and/or the hydrocarbyl substituent of C1 position.

Shown in reaction scheme 6, at first in the A of steroide ring, produce 1-alkene-3-ketone functional group, by adopting in the multiple alkoxide anion any to carry out Michael reaction, the oxygen functional group is introduced in the C1 position that can be implemented in the steroide carbon skeleton again.For example, ketenes 29 can adopt standard method by compound 13 preparations.Benzyloxy compound 30 can be produced by making ketenes 29 and benzylalcohol and potassium hydroxide reaction.With the C3 ketone reduction of compound 30, the secondary alcohol that forms is protected (obtaining compound 31) with the siloxy-derivative, carry out catalytic hydrogenation again, obtain the compound 32 of C1 hydroxyl functional base.Re-use for example CH of PDC ₂Cl ₂Solution carries out oxidation to this secondary alcohol, produces the compound 33 with C1 ketone.

Reaction scheme 6

The compound that comprises alkyl group and oh group simultaneously in the C1 position can obtain by making compound 33 and alkyl lithium reagents reaction.For example, compound 33 will obtain the tertiary alcohol, compound 34 (reaction scheme 7) with the ethereal solution reaction of lithium methide.

Reaction scheme 7

Michael reaction and reaction scheme 6 described processes are similar, can be used for adding alkyl to the C1 position.This process can comprise R by use ₂CuLi (R can be alkyl, vinyl or aryl) finishes in interior plurality of reagents.For example, compound 29 can with Me ₂The ethereal solution reaction of CuLi obtains the methyl substituted derivative 35 of C1 (reaction scheme 8).

Reaction scheme 8

The compound of structural formula 6 can have oxygen and/or hydrocarbyl substituent in the C2 position.The C2 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of oxygen and/or hydrocarbyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 compound of C2 position aerobic and/or hydrocarbyl substituent provides the oxygen and/or the hydrocarbyl substituent of C2 position.

Wrapping oxygen containing compound in the C2 position can prepare in many ways, comprises the hydroboration of the silyl enol ether shown in reaction scheme 9.The silyl enol ether can re-use TMSCl the enolate that forms is caught by ketenes 29 through lithium/ammonia reduction preparation, obtains compound 36 (or other R ₃SiCl reagent is to produce similar silyl enol ether).The hydrogen boronation reaction of the carbon-to-carbon double bond in 36 can provide 2 α, 3 beta-dihydroxyl functionalized form (compound 19).Use the CH of PDC ₂Cl ₂Solution carries out oxidation to this dihydroxy compound can obtain diketone 38.

Reaction scheme 9

For example can carry out the alpha-alkyl reaction, produce the compound that the C2 hydrocarbon replaces by making the compound that comprises C3 ketone functional group.For example,, with alkylating reagent the negatively charged ion that forms is caught again, C22 is provided alkylation with lithium/ammonia reduction ketenes 29.Handle the enolate that forms with methyl-iodide and can obtain the methylated compound 39 of C2 (following reaction scheme 10).This method can adopt many different alkylogens to be used for various compound.

Reaction scheme 10

The compound of structural formula 6 can have hydrocarbyl substituent in the C3 position.The C3 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of hydrocarbyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 C3 position has the compound of hydrocarbyl substituent that the hydrocarbyl substituent of C3 position is provided.

Compound 13 is carried out Wittig reaction, again with the reduction of two keys or carry out other modification and will be provided at the derivative that the C3 position has alkyl or dialkyl group.For example, compound 13 can be used for preparing compound 40 (reaction scheme 11) with the toluene solution reaction of first base three phenyl phosphonium bromides and potassium tert.-butoxide.With methylene iodide and zinc-copper compound is carried out the Simmons-Smith reaction, the Pd/C that re-uses in hydrogen and the ethanol carries out catalytic hydrogenolysis to cyclopropane derivative 41, obtains dialkyl derivatives 42 (reaction scheme 11).

Reaction scheme 11

The compound of structural formula 6 can have hydrocarbyl substituent in the C4 position.The C4 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of hydrocarbyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 C4 position has the compound of hydrocarbyl substituent that the hydrocarbyl substituent of C4 position is provided.

At first produce enolate negatively charged ion (for example using the lithium in the liquefied ammonia), handle with suitable alkylogen again, can obtain the C4 alkylation, shown in reaction scheme 12 by compound 10 ketenes.

Reaction scheme 12

In addition, the compound of structural formula 6 can have carbonyl functional group in the C4 position.The C4 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of carbonyl functional group.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 C4 position has the compound of carbonyl functional group that the carbonyl functional group of C4 position is provided.As described below, the carbonyl functional group in the C4 position provides the approach that makes things convenient for that synthesizes the compound that has the tertiary alcohol and alkyl in the C4 position.

The compound that has ketone (carbonyl) functional group in the C4 position can be by compound 44 preparation (it again can from reaction scheme 44 by the preparation of acetic ester deacetylation), and the oxidizing reaction of the 4 beta-hydroxy functional groups that preparation process can be by selectivity silylation reactive, epoxidation reaction and epoxide ring-opening reaction and formation is finished.For example, shown in reaction scheme 13, the Tosyl chloride that glycol 44 is used among pyridine and the DMF is handled, and can introduce 3 β, 4 beta epoxide things (compound 46) with silylanizing thing 45 reactions of potassium tert.-butoxide and formation again.Use Me ₂CuLi handles epoxide, obtains 3 Alpha-Methyl derivatives 47, subsequently, uses for example CH of PDC again ₂Cl ₂Solution carries out oxidation, obtains the required ketone in the C4 position (carbonyl) (compound 48).Use the t-butanol solution of potassium tert.-butoxide to use the THF solution-treated ketone of lithium methide again, obtain the tertiary alcohol (compound 49) of C4 position the epimerization of 3 Beta-methyl derivatives.

Reaction scheme 13

In addition, the compound of structural formula 6 can have oxygen or hydrocarbyl substituent in the C5 position.The C5 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of oxygen or hydrocarbyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 compound of C5 position aerobic or hydrocarbyl substituent provides the oxygen or the hydrocarbyl substituent of C5 position.

Compound 10 is carried out epoxidation reaction, produce hydroxyl by open loop again, the alkoxyl group that carries out the C5 position of carbon skeleton subsequently replaces.For example, the epoxidation of two keys can produce corresponding epoxide derivate 50 in the compound 10, and this derivative is easy to transform into tert-hydroxyl compound 24 (following reaction scheme 14).Subsequently, the THF solution that uses sodium borohydride carries out methylation reaction with methyl-iodide with compound 24 reduction in the presence of the THF of potassium tert.-butoxide solution, can provide diacetoxy compound 51 (following reaction scheme 15).Alkyl replacement in the C5 position can use suitable alkyl copper lithium reagent to finish.For example, use Me ₂The diethyl ether solution of CuLi is handled compound 10 can produce C5 methyl-derivatives 52 (reaction scheme 16).

Reaction scheme 14

Reaction scheme 15

Reaction scheme 16

The compound of structural formula 6 can have oxygen and/or hydrocarbyl substituent in the C9 position.The C9 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of oxygen or hydrocarbyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 compound of C9 position aerobic or hydrocarbyl substituent provides the oxygen or the hydrocarbyl substituent of C9 position.

Shown in reaction scheme 17, pass through Δ ^9.11LiAlH is used in olefin(e) compound and metachloroperbenzoic acid reaction again ₄Reduce, can realize the hydroxylation of C9 position.For example, utilize this process, compound 53 (is made by compound 60 hydrogenations; For example use sodium hydride, dithiocarbonic anhydride, methyl-iodide, and heat) can be used as the raw material of production compound 54, compound 54 comprises C9 position hydroxy derivative 55 through epoxide is reduced to produce.After, the tertiary alcohol in the compound 55 reacts with the methyl-sulfate in aqueous sodium hydroxide solution, can obtain corresponding alkoxy derivative, compound 56.

Reaction scheme 17

In addition, the compound of structural formula 6 can have hydrocarbyl substituent in the C9 position.The C9 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of hydrocarbyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 C9 position has the compound of hydrocarbyl substituent that the hydrocarbyl substituent of C9 position is provided.

Adopt methylene iodide and zinc-copper to make compound Cyclopropanated, carry out catalytic hydrogenation again, can obtain the compound 57 (reaction scheme 18) that corresponding C 9-alkyl replaces.

Reaction scheme 18

In addition, the compound of structural formula 6 can have halogenic substituent in the C9 position.The C9 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of halogenic substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 C9 position has the compound of halogenic substituent that the halogenic substituent of C9 position is provided.

Can adopt multiple mode to realize introducing halogen atom, comprise making the C9 tertiary alcohol (referring to, the compound 55 in the reaction scheme 17 for example) and thionyl chloride reaction to the C9 position.For example, compound 55 and CH ₂C1 ₂In thionyl chloride reaction can be used for providing chlorinated derivative 59, shown in reaction scheme 19.

Reaction scheme 19

The compound of structural formula 6 preferably has methyl substituents in the C10 position.But can be derived in the C10 position, to have the many functional groups except that methyl.The C10 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of oxygen or hydrocarbyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 compound of C10 position aerobic or hydrocarbyl substituent provides the oxygen or the hydrocarbyl substituent of C10 position.

The derivatization reaction of C10 position can be realized through the path shown in the reaction scheme 20.Adopt the pyridine solution of nitrosyl chloride (NOCl) that 10 beta-hydroxy steroides 60 (its preparation example is shown in following reaction scheme 22) are derived, obtain a kind of nitrous acid derivative, as 61.This nitrous acid derivative 61 is carried out the mixture that irradiation can obtain oxime 62 and 63.Han Shui dioxane solution and acetic acid treatment by with titanous chloride can be reduced into compound 63 corresponding imines 64.Adopt the acetic acid aqueous solution of Sodium Nitrite that compound 64 is handled, can produce hemiacetal acetate compound 65.This also can cause 6, the deprotection of 7-oh group.Make crude product and 2,2-Propanal dimethyl acetal and camphorsulfonic acid reaction can be introduced acetone solvate again.By alkaline hydrolysis (sodium hydroxide, methyl alcohol), obtain hydroxy aldehyde 66, adopt the DMF solution of BnBr, NaH that C11 position secondary alcohol is protected as benzylic ether again, can obtain compound 67.

Reaction scheme 20

Compound 67 and CH ₃MgBr carries out Grignard reaction, passes through at CH again ₂Cl ₂In carry out the PDC oxidation, use the CH of metachloroperbenzoic acid again ₂Cl ₂Solution carries out the Bayer-williger oxidizing reaction, obtains C10 acetoxyl group derivative 68.Adopt the methanol solution of alkali such as sodium methylate to remove aceticoceptor, obtain C10-β alcohol 69.Then, again the hydroxyl of C10 is further derived, for example use the THF solution of sodium hydride, handle with alkylating agent such as methyl-iodide again, obtain alkoxy compound 70.In addition, can adopt chlorizating agent such as thionyl chloride that the C10 oh group in the compound 69 is changed into corresponding chlorinated thing derivative 71, shown in reaction scheme 21.

Reaction scheme 21

The compound of structural formula 6 has oxygen and/or hydrocarbyl substituent in the C11 position.The C11 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of oxygen and/or hydrocarbyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 compound of C11 position aerobic or hydrocarbyl substituent provides the oxygen or the hydrocarbyl substituent of C11 position.

Comprise the preparation process of structural formula 6 compounds of C11 position oxygen functional group can be shown in reaction scheme 22 path, finish by commercially available raw material 72 and related compound.

Reaction scheme 22

If being used in the steroide that has hydroxyl on the A ring is raw material, as compound 75 (it can prepare (reaction scheme 22) by commercial compound 72), adopt two step processes can remove the hydroxyl of C3 position, described two step processes comprise the THF formulations prepared from solutions methyl xanthate that adopts sodium hydride, dithiocarbonic anhydride and methyl-iodide, use nBu again ₃SnH reduces, and deprotection (80% acetate) obtains compound 77.Adopt the methanol solution of sodium borohydride, after with the DMF solution of TBDMSCl and imidazoles C17 ketone being reduced and protects again, can adopt the CH of many kinds of oxidizing conditions such as chromium trioxide and 3 ₂Cl ₂The water of solution or ruthenium trichloride and tBuOOH and cyclohexane solution are realized the oxidation of C7 position.Then, ketone reduction (sodium borohydride, cerous compounds, THF-methyl alcohol) and acetylize with the C7 position obtain C7 acetoxyl group derivative 80.With compound 80 hydroborations, obtain having 6 α, 7 β, the product of 11 beta-hydroxy forms, promptly triol 81.In the presence of camphorsulfonic acid (CSA), adopt 2, the 2-Propanal dimethyl acetal is to 6 α of compound 81, and 7 β hydroxyls are protected, and use the CH of PDC again ₂Cl ₂Solution carries out oxidation, obtains comprising the compound 82 of C11 ketone.

The compound of structural formula 6 also can have hydrocarbyl substituent in the C11 position.The C11 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of hydrocarbyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 C11 position has the compound of hydrocarbyl substituent that the hydrocarbyl substituent of C11 position is provided.

Can C11 ketone group steroide 82 be changed into corresponding tetraalkyl center according to process shown in the reaction scheme 23.

Reaction scheme 23

The toluene solution of C11 ketone group steroide 82 can be added in the solution of methyltriphenylphospbromide bromide and potassium tert.-butoxide, obtain having Δ ¹¹The compound 83 of carbon-to-carbon double bond.Handle compound 83 with methylene iodide, zinc-copper and can obtain cyclopropyl derivatives 84.With cyclopropane ring hydrogenation (hydrogen, Pd/carbon catalyst, ethanol), obtain dialkyl derivatives 85.Also can adopt other Witting reagent to prepare the steroide that similar alkyl replaces.

The monoalkylation reaction of C11 position can directly be carried out catalytic hydrogenation and finish (as reaction scheme 24) by as previously mentioned C11 ketone being carried out Wittig reaction again.For example, the catalytic hydrogenation (hydrogen, Pd/carbon catalyst, ethanol) to compound 83 obtains C11 methylated steroids 86.

Reaction scheme 24

The compound of structural formula 6 can have halogenic substituent in the C11 position.The C11 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of halogenic substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 C11 position has the compound of halogenic substituent that the halogenic substituent of C11 position is provided.

The halogenating reaction of C11 position can be carried out according to the approach shown in the reaction scheme 25.For example, use the CH of halogenating agent such as thionyl chloride ₂Cl ₂Solution carries out halogenation to compound 60, obtains corresponding 11 β-chlorine derivative 87.Usually, the hydroxyl functional base can be used as the precursor that forms the halogen functional group.

Reaction scheme 25

The compound of structural formula 6 can have oxygen and/or hydrocarbyl substituent in the C12 position.The C12 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of oxygen and/or hydrocarbyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 compound of C12 position aerobic or hydrocarbyl substituent provides the oxygen or the hydrocarbyl substituent of C12 position.

The process that the oxygen functional group is provided in the C12 position is shown in reaction scheme 26.

Reaction scheme 26

C11 ketone group steroide such as compound 82 can with the THF solution reaction of LDA, use (Me again ₃N) ₂P (O) Cl catches enolate anion, adopts lithium and ethamine that the phosphoric acid enol is reduced again, obtains having Δ ^11,12The compound of carbon-to-carbon double bond such as compound 88.Adopt the CH of epoxidizing agent such as mCPBA ₂Cl ₂Solution carries out epoxidation reaction, obtains corresponding 11 α, 12 α-epoxidized derivs 89.After, use LiAlH ₄Epoxide is reduced, obtain corresponding 12 Alpha-hydroxy derivatives 90, adopt the suitable oxygenant such as the CH of dichromic acid pyridine (PDC) ₂Cl ₂Solution obtains required C12 ketone group steroide 91 with derivative 90 oxidations.

The compound of structural formula 6 can have hydrocarbyl substituent in the C12 position.The C12 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of hydrocarbyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 C12 position has the compound of hydrocarbyl substituent that the hydrocarbyl substituent of C12 position is provided.

Can on the C12 position, introduce alkyl such as methyl according to reaction scheme as follows 27.C11 ketone group steroide 82 (as according to reaction scheme 22 preparations) merges with the THF solution of highly basic such as di-isopropyl lithamide, handles with alkylating agent such as methyl-iodide, obtains C12 alkylate 92.In this step, C11 ketone can adopt several different methods to remove, comprise reaction scheme 26 described those, thereby obtain monomethylation product 93.Further handle, obtain C12 dimethyl product 94 with highly basic such as di-isopropyl lithamide and with alkylating agent such as methyl-iodide.Equally, this compound can be placed under the reductive condition, remove the C11 ketone group, thereby obtain C12 dimethyl derivative 95.

Reaction scheme 27

The compound of structural formula 6 can have oxygen and hydrocarbyl substituent in the C12 position.The C12 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of oxygen and hydrocarbyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 compound of C12 position aerobic and hydrocarbyl substituent provides the oxygen and the hydrocarbyl substituent of C12 position.

Reaction scheme 28 has provided the method that is prepared the C12 position tertiary alcohol by corresponding C 12 ketone.In reaction scheme 28, the diethyl ether solution of C12 ketone 91 usefulness alkyl lithium reagents such as lithium methide is handled, and obtains the required tertiary alcohol 96.

Reaction scheme 28

The compound of structural formula 6 can have carbon, oxygen or the halogen that links to each other with C13, only mentions the minority atom here.The C13 position that below is given in structural formula 6 compounds provides this substituent illustrative synthetic method.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 C13 position has this substituent compound that the identical or similar substituting group of C13 position is provided.

Substituting group in the C13 position can be introduced according to following reaction scheme 29.With with previous reaction route 20 similar modes, available-oxyl in C13 position such as methoxyl group replace.For example, adopt the moisture dioxane solution and the acetate of titanous chloride to handle, 9 oxime derivate 62 (as according to reaction scheme 20 preparations) can be reduced into corresponding imines 97.Aqueous acetic acid solution-treated compound 97 with Sodium Nitrite can obtain hemiacetal acetate compound 98.By alkaline hydrolysis (sodium hydroxide, methyl alcohol), obtain hydroxy aldehyde 99, adopt the DMF solution of BnBr, NaH that the secondary alcohol of C11 position is protected as benzylic ether again, can obtain compound 100.

Reaction scheme 29

Can carry out Grignard reaction on C13, to introduce another functional group to compound 100.For example, use CH ₃MgBr handles compound 100, again the secondary alcohol that forms is carried out oxidation, obtains the methyl ketone substituting group of C13 position.It is used the CH of metachloroperbenzoic acid again ₂Cl ₂Solution carries out the Bayer-Williger oxidizing reaction, obtains C13 acetoxyl group derivative 101.Adopt the methanol solution of alkali such as sodium methylate that this ester is handled, obtain the tertiary alcohol 102.Then, alcohol reacts with the THF solution of sodium hydride, handles with methyl-iodide, produces C13 methoxyl group steroide 103.Other alkylating agent can be used for preparing other-oxyl derivative.By alcohol 102 and thionyl chloride reaction, change into halogenide such as muriate, thereby obtain C13 chlorine steroide 104, shown in reaction scheme 30.

Reaction scheme 30

The compound of structural formula 6 can have hydrocarbyl substituent in the C14 position.The C14 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of hydrocarbyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 C14 position has the compound of hydrocarbyl substituent that the hydrocarbyl substituent of C14 position is provided.

For example, introducing alkyl to the C14 position of steroide carbon skeleton can finish by the alkylated reaction of C14 position.A kind of method is to adopt the alkylated reaction shown in the reaction scheme 31.During beginning, can use the CH of PDC again by compound 105 being carried out deprotection (TBAF, THF) ₂Cl ₂Solution carries out oxidation to secondary alcohol, obtains C17 ketone derivatives 106, refabrication ketenes 107.Adopt acetate pseudoallyl ester and pTsOH to produce intermediate enol acetic ester, reagent shown in the reaction scheme 31 produces ketenes for another example, can realize ketone 106 is converted into ketenes 107.After this, again by making the THF solution reaction of ketenes 107 and diethyl ammonification lithium, with the negatively charged ion and trifluoromethanesulfonic acid triisopropyl silyl ester (TIPSOTf) reaction that form, ketenes 107 is changed into silyl enol ether 108 again.Then, adopt methylene iodide and zinc-copper by silyl ether 108 preparation cyclopropane derivatives 109.Adopt the THF solution of TBAF, DMSO solution and the moisture last handling process by tBuOK carries out silyl enol ether deprotection and makes the cyclopropane ring cracking again.

Reaction scheme 31

The compound of structural formula 6 can have oxygen and/or hydrocarbyl substituent in the C15 position.The C15 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of oxygen and/or hydrocarbyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 compound of C15 position aerobic or hydrocarbyl substituent provides the oxygen or the hydrocarbyl substituent of C15 position.

For example, adopt any alcoxyl negatively charged ion to finish the introducing of oxygen functional group on the C15 position of steroide carbon skeleton by the reaction of Michael addition type.Shown in reaction scheme 32; (be the representational C15--oxyl of the present invention steroid derivatives by making ketenes 107 can produce 4-methoxybenzyl oxycompound 111 with 4-methoxy-benzyl alcohol and alkali (as powder KOH) reaction; wherein, 4-methoxyl group benzyloxy base (MPMO) is as hydroxy-protective group).4-methoxy-benzyl protecting group can be removed under oxidizing condition, for example adopts 2,3-two chloro-5, and 6-dicyano-1,4-benzoquinones (DDQ) carries out oxidation, obtains C15 hydroxyl (compound 112).When this process rear oxidation secondary alcohol (for example uses the CH of PDC ₂Cl ₂Solution) time, can produce corresponding C 15 ketone (compound 113).

Reaction scheme 32

The compound that comprises alkyl in the C15 position also can be produced by Michael's type conjugate addition reaction.For example, make the sour lithium of compound 107 and organic copper (as Me ₂CuLi) diethyl ether solution reaction is used for producing methyl-derivatives 114, shown in reaction scheme 33.

Reaction scheme 33

The compound that had not only comprised alkyl (as alkyl) but also comprised-oxyl (as alkoxyl group) on the C15 position can produce by compound 117 is carried out Grignard reaction, shown in reaction scheme 34.Compound 117 prepares with three step reaction, and described step comprises the methyl xanthate nBu by 111 preparations of C17 hydroxyl similar compound ₃SnH reduces and obtains steroide 115, reoxidizes to remove (for example using DDQ) MPM protecting group to produce secondary alcohol derivative 116.Subsequently, compound 116 is oxidized to corresponding ketone compound 117.Adopt alkyl bromination azoviolet (as CH ₃MgBr) ethereal solution carries out Grignard reaction to compound 117, produces the tertiary alcohol 118.In the presence of alkali (as salt of wormwood); ((note: at reaction scheme 34 and have herein in all reaction schemes of alkylating agent with alkylating agent as methyl-iodide; also available acylating agent replaces alkylating agent)) tertiary alcohol in 118 is carried out methylation reaction, obtain uncle's methoxylation compound 119.

Reaction scheme 34

The compound of structural formula 6 can have oxygen and/or hydrocarbyl substituent in the C16 position.The C16 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of oxygen and/or hydrocarbyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 compound of C16 position aerobic or hydrocarbyl substituent provides the oxygen or the hydrocarbyl substituent of C16 position.

Shown in reaction scheme 35, compound 121 is carried out Grignard reaction, can realize the introducing of the C16 position tert-hydroxyl of steroide carbon skeleton.Ketone 121 can (for example use Sia by compound 308 (being prepared by compound 106 as reaction scheme 35) is carried out olefin hydrogenation ₃The THF solution of BH is used sodium hydroxide, hydrogen peroxide again) produce.Then, for example use the CH of PDC ₂Cl ₂Solution carries out oxidation to the secondary alcohol on the C16 and produces required C16 ketone functional group, obtains compound 121.With ketone 121 usefulness Grignard reagent such as CH ₃The diethyl ether solution reduction of MgBr is used for producing corresponding tertiary alcohol derivative, is compound 122 in this example.Corresponding alkoxy derivative 123 can directly use suitable alkali and alkylogen to produce by compound 122.

Reaction scheme 35

The alkoxyl group of C16 position can be directly by corresponding hydroxy compound deposits yields.For example, compound 124 can produce by making compound 120 and reagent react as methyl-iodide and alkali such as salt of wormwood, sees reaction scheme 36.

Reaction scheme 36

The C16 alkyl can be introduced by the compound that comprises the C17 carbonyl is carried out direct alkylated reaction.For example, the THF solution reaction of compound 106 and methyl-iodide and LDA (other highly basic and alkylating agent also can use) produces the compound 125 of C16 methyl, shown in reaction scheme 37.

Reaction scheme 37

The compound of structural formula 6 can have oxygen and/or hydrocarbyl substituent in the C17 position, comprise the tertiary alcohol and hydroxyl functional base.The C17 position that below is given in structural formula 6 compounds provides the illustrative synthetic method of the tertiary alcohol and hydroxyl substituent.Should be appreciated that and to adopt identical or similar synthetic method to wish that to any of structural formula 5-12 C17 position has the compound of the tertiary alcohol and hydroxyl substituent that the tertiary alcohol and the hydroxyl substituent of C17 position are provided.

Can be used on the C17 position, adding tertiary alcohol functional group with reaction scheme 34 similar Grignard reactions.For example, shown in reaction scheme 38, compound 106 can be directly and CH ₃The diethyl ether solution reaction of MgBr produces tertiary alcohol derivative 126.The tertiary alcohol that forms is methylated, obtain corresponding C 17 methoxylation compounds 127.Certainly, other alkylating agent also is used to provide various hydrocarbon oxy compounds.

Reaction scheme 38

One aspect of the present invention provides in C6 and C7 position hydroxylation, has had 5 α hydrogen and C3 and be not connected the steroide that the C5 steric configuration of Sauerstoffatom is determined.In one embodiment, the compounds of group that this steric configuration is determined has structural formula 7, comprises enantiomorph or geometrical isomer that it is independent, further comprises its solvate or pharmacy acceptable salt.Structural formula 7 is defined as follows:

The compound of following formula

Comprise its pharmacy acceptable salt and its solvate, wherein:

C3 quilt=C (R ⁴) (R ⁴) and-C (R ⁴) (R ⁴) (C (R ⁴(R ⁴)) _n-replace, wherein n is 1 to about 6, perhaps by-X and-R ⁴In two replacements, condition is that C3 does not link to each other with Sauerstoffatom;

R ¹Be H, thereby a kind of blocking group makes-OR ¹Be the oh group of protection, wherein, ortho position-OR ¹Group can form the ring structure of protection vicinal hydroxyl groups together, and geminal-OR ¹Group can form the ring structure of protection carbonyl together, and condition is one-OR on C6 and C7 position ¹Or two-OR ¹Represent the carbonyl of carbonyl or protection;

Each R ⁴Be independently selected from H and C ₁-C ₃₀Organic residue, this organic residue can optionally comprise at least one heteroatoms, and described heteroatoms is selected from boron, halogen, nitrogen, oxygen, silicon and sulphur, wherein, two geminal R ⁴Group can form a ring with the carbon atom that is connected with them; With

X represents fluorine, chlorine, bromine and iodine.

The compound of structural formula 7 has hydroxyl and has 5C hydrogen at C6 and C7 place.In reaction scheme 2, proposed to be used to prepare the synthetic route of compound, shown the preparation process of compound 11 with these constitutional featuress.Because this compound has the Sauerstoffatom that links to each other with C3, thereby compound 11 is not the representative compounds of structural formula 7, compound 11 can be changed into the compound of structural formula 7.Shown in reaction scheme 39, the lithium/trimethyl carbinol in the liquefied ammonia can be used for obtaining required reduzate.The hydrogenation of compound 128 can provide compound 105, and it has-CH in the C3 position ₂-group is seen reaction scheme 39 equally.

Reaction scheme 39

Shown in reaction scheme 40, also compound 128 can be changed into the compound of another kind of structural formula 7.For example, the hydroxyl deprotection that the C17 of compound 128 can be protected is to produce compound 129, and then, the C17 hydroxyl oxygen with compound 129 changes into the C17 carbonyl again, shown in compound 106.

Reaction scheme 40

The compound that comprises the structural formula 7 of methylene radical in the C3 position can be made by the hydroxyl that has hydroxyl, protection in the C3 position or the compound of ketone functional group.Can adopt identical or similar synthetic method to prepare the compound that C3 position wherein needs any structural formula 5-12 of methylene radical.

For example, above-mentioned reaction scheme 39 has been described to adopt and has been reacted the method that compound 11 is changed into compound 105 as previously mentioned.Therefore, each reaction that interrelates with each reaction scheme of the present invention can extend to and comprise and variously contain methylene radical and the compound of non-hydroxyl or carbonyl in the C3 position.But, in some cases, at first require a series of protections and/or deprotection steps.

Reaction scheme 41 has shown an example, and wherein, C3 siloxy-functional group must at first carry out deprotection before deoxygenation.TBDMS group in compound 31 can adopt the THF solution of TBAF to remove.Using potassium hydride KH, dithiocarbonic anhydride and methyl-iodide to prepare the methyl xanthate derivative is compound 130, passes through nBu again ₃SnH reduction obtains the compound 131 that comprises methylene radical in the C3 position and comprise the hydroxyl of protection in the C1 position.Then, by removing the C1 protecting group, re-use suitable oxygenant such as the CH of PDC ₂Cl ₂The oxidable one-tenth of solution C1 ketone obtains compound 132.

Reaction scheme 41

Have C3 alkyl functional base, comprise that the compound of the structural formula 5-12 of structural formula 7 can obtain (being prepared by C3 ketone) by Wittig reaction as described in reaction scheme 11.Various Witting reagents can be used for this purpose, thereby produce the substituting group of various chain lengths and side chain situation.

One aspect of the present invention provides at C6 and C7 position to have oxygen and/or hydrocarbyl substituent, but at C10 and C13 and not all have the demethylation steroide of methyl.In one embodiment, the demethylation steroide has structural formula 8, comprises enantiomorph or geometrical isomer that it is independent, further comprises its solvate or pharmacy acceptable salt.Structural formula 8 is defined as follows:

The compound of following formula

Comprise its pharmacy acceptable salt and its solvate, wherein:

Condition is: (a) C10 and C13 do not have methyl substituents and (b) as C10 during by methyl substituted, then C14 is not by methyl substituted simultaneously;

Each R ⁴All be independently selected from H and C ₁-C ₃₀Organic residue, this organic residue can optionally comprise at least one heteroatoms, and described heteroatoms is by boron, halogen, nitrogen, oxygen, silicon and sulphur, wherein, two geminal R ⁴Group can form a ring with the carbon atom that is connected with them; With

X represents fluorine, chlorine, bromine and iodine.

The more substituent examples of compounds that comprise except that methyl at C10 or C13 are described in conjunction with reaction scheme 20,29 and 30 herein.Substituting group comprises carbonyl, hydroxyl methylene radical, methoxyl group, ketal, interior ester carbonyl group, aldehyde, hydroxyl etc.The compound that does not comprise substituting group (promptly only being replaced by hydrogen) in C10 and/or C13 position is described in reaction scheme 20,29 and 30.Below, generation 19-removes first-6 α, the example of the synthetic method of 7 β-titanium dioxide steroide for being discussed.

The synthetic method of the various compound of many present invention binding compounds 1 and 247 is described in detail, and compound 1 and 247 is commercially available raw material.But similar compound is as only being that the preparation process that lacks the compound 141 of C10 methyl substituents can be finished according to following reaction scheme 42.

In reaction scheme 42, raw material be commercially available 19-go first-testosterone 133 (Steraloids Inc., Wilton, NH or Aldrich Chemical Company, Milwaukee, WI).Adopt the ethanolic soln of sodium borohydride that compound 133 is reduced, obtain comprising the compound 134 of 3 beta-hydroxies.After the DMF solution that adopts TBDMSCl and imidazoles is protected 3 beta-hydroxies, the two protection compounds 135 that form are carried out the allylic oxidation, obtain ketenes derivative 136.After preamble (reaction scheme 1) described reduction and acetylize, adopt BH again ₃-THF carries out hydroboration and oxidation processes (hydrogen peroxide, 30% sodium hydroxide), obtains comprising 6 α, 7 β, the compound 138 of 17 beta-hydroxy forms.Adopt 2,2-Propanal dimethyl acetal and camphorsulfonic acid are to 6 α, and 7 beta-hydroxies are protected, and use the CH of PDC again ₂Cl ₂Solution carries out oxidation to the C17 hydroxyl, obtains comprising the compound 140 of C17 ketone functional group.Compound 140 and obtain methylene derivatives by the Witting reagent reaction of the toluene solution of ethyl triphenyl bromination and tBuOK preparation; its can be in acetate deprotection; obtain trihydroxy-compound 141, it is identical with compound 333, just lacks the C10 methyl substituents.

Reaction scheme 42

One aspect of the present invention, provide on C3, C4, C6 and each position, C7 position all to have oxygen and/or hydrocarbyl substituent, wherein oxygen on the C6 position and/or hydrocarbyl substituent have α stereoisomerism and oxygen and/or hydrocarbyl substituent on the C7 position has the stereomeric polyoxy steroide of β.In one embodiment, this polyoxy steroide has structural formula 9, comprises enantiomorph or geometrical isomer that it is independent, further comprises its solvate or pharmacy acceptable salt.Structural formula 9 is defined as follows:

Compound with following formula

Compound also comprises its pharmacy acceptable salt and solvate, wherein:

(b) two in the following formula, each is independently selected from :-X ,-R ⁴With-OR ¹Condition is that C17 states any replacement in the group:

C8 quilt-X or-R ⁴Replace, preferably directly be not connected with oxygen;

R ¹Be H, thereby a kind of blocking group makes-OR ¹Be the oh group of protection, wherein, ortho position-OR ¹Group can form the ring structure of protection vicinal hydroxyl groups together, and geminal-OR ¹Group can form the ring structure of protection carbonyl together;

X represents fluorine, chlorine, bromine and iodine.

The compound with oxygen and/or hydrocarbyl substituent shown in the structural formula 9 can be prepared by compound 142, and compound 142 can be according to following reaction scheme 52 described processes preparations.As described in reaction scheme 43, compound 142 can carry out epoxidation with any epoxidation conditions to be handled, as adopting the CH of metachloroperbenzoic acid ₂Cl ₂Solution obtains epoxidation compound 143.Weak organic acid (as anhydrous acetic acid, it also is preferred acid) is adopted in the open loop of epoxide, obtains compound 144, and it is the representative compounds of structural formula 9.

Reaction scheme 43

From compound 144, can prepare the compound of multiple other structural formula 9.For example, shown in reaction scheme 43, can compound 144 is deacetylated to obtain tetrahydroxy ketone compound 145.Ketone group on the C17 position can carry out foregoing Wittig reaction, obtains the tetrahydroxy olefin(e) compound of a big class formation formula 9.

Have 3,4,6, the structural formula 9 of 7-four oxidised forms also can comprise oxygen-containing substituents.For example, the compound of structural formula 9 can have a Sauerstoffatom at place, C11 position.The synthetic method of introducing the C11 Sauerstoffatom can be by finishing as reaction scheme 44 or by finishing with the similar reaction of reaction scheme 44, this synthetic method can be used for preparing the compound of the structural formula 5-12 that comprises structural formula 9.

For example, do not use commercially available have C11 hydroxyl functional base or Δ ^9,11The raw material of carbon-to-carbon double bond, and between forming, produce required behind the dichloroiodo benzyl carbamoyl ester at position △ by photodissociation ^9,11Unsaturated link(age) (compound 149).C6 in the compound 146 and C7 hydroxyl (according to reaction scheme 61 preparations) can adopt 2, and 2-Propanal dimethyl acetal and camphorsulfonic acid are protected to obtain compound 147.Subsequently, the reaction of the pyridine solution of a compound 147 and a dichloroiodo benzyl formyl chloride obtains compound 149 by carrying out photolysis in the tetracol phenixin again.Obtain C11 hydroxy derivatives 151 by hydroboration/oxidation behind the hydroxyl of protection A ring.Adopt 80% acetate to carry out complete deprotection and obtain six alcohol 152.

Reaction scheme 44

One aspect of the present invention provides the steroid ketone compound that has pyrans or δ lactonic ring on the C17 side chain.In one embodiment, this steroid ketone compound has structural formula 10, comprises enantiomorph or geometrical isomer that it is independent, further comprises its solvate or pharmacy acceptable salt.Structural formula 10 is defined as follows:

Compound with following formula Comprise its pharmacy acceptable salt and solvate, wherein:

C1, C2, C3, C4, C6, C7, C11, C12, C15 and C16 are replaced by following radicals independently of one another:

Condition is that C3, C4, C6 and C7 are not replaced by the hydroxyl of hydroxyl or protection simultaneously, are not preferably replaced by Sauerstoffatom simultaneously;

G is-C (=O)-,-CH (OR ¹)-,-C (R ⁴) (OR ¹)-or-C (OR ¹) (OR ¹)-;

X represents fluorine, chlorine, bromine and iodine.

The method that makes things convenient for of introducing the C17 side chain in structural formula 10 compounds is from the L-Karvon, shown in reaction scheme 45.

Reaction scheme 45

L-Karvon (153) can change into compound 154 according to the method that provides in the document, for example, referring to: tetrahedron communication (Tetrahedron Letters) 25 (41): 4685-4688 (1984).Then, by for example being converted into acetic ester the primary alconol in the compound 154 is protected.The employing acidic conditions is removed the ketal protected group in the compound 155, obtains aldehyde 156.

Shown in reaction scheme 46, compound 156 can provide the C17 side chain of structural formula 10 compounds.Compound 145 as reaction scheme 43 preparations can be handled with ylide, obtains compound 157 (as the raw material of reaction scheme 46), and described ylide is by ethyl triphenyl phosphonium bromide and alkali preparation.Then, four oh groups can be converted to the oh group of protection, and benzyloxy for example is shown in compound 158.Then, in the presence of Lewis acid, make compound 158 and aldehyde 156 (reaction scheme 45) coupling, obtain compound 159.After this, with alkali the acetoxyl group of C29 is carried out deprotection, obtain diol compound 160, compound 160 can be oxidized to delta-lactone compound 161.Compound 161 is carried out the allylic oxidation can be oxidized to benzyl (Bn) benzoic ether (Bz) group simultaneously at C15 position carbonylate, forms compound 162.

With the conjugated Δ in the D ring of compound 162 ¹⁶The carbon-to-carbon double bond reduction obtains compound 163.With alkaline condition (for example, the methanol solution of sodium methylate) removes benzoic ether group in 163, simultaneously C14 is carried out epimerization, obtain product 164, this product comprises the epimer mixture that contains the compound that is connected with anti-C/D ring along the connection of C/D ring.At last, the ketone of protection C15 changes into lactonaphthol with delta-lactone again; and deprotection (80% acetate), obtain 22,29-epoxy-3; 4,6,7; the epimer 22 of 29-penta hydroxy group-14 β-stigmastane-15-ketone (compound 165) and its C14 position; 29-epoxy-3,4,6; 7,29-penta hydroxy group-14 α-stigmastane-15-ketone.

Reaction scheme 46

The compound of structural formula 10 can have C15 ketone and C22,29 epoxide function bases.In fact, can adopt following combined method to produce except that C15 ketone and side chain hemiacetal, in the A-D ring, contain the compound of various functional groups.

For example, shown in reaction scheme 47, can use method as described below to be created in the C3 position and contain methylene radical, contain carbonyl and contain the compound 176 of side chain hemiacetal in the C15 position.C15 ketone and side chain hemiacetal can adopt the method that as above describes in detail to introduce (association reaction route 45 and 46).

Reaction scheme 47

Shown in reaction scheme 47, compound 76 can adopt the palladium/charcoal in hydrogen, the ethanol to carry out deprotection, to obtain comprising the compound of C11 hydroxyl functional base, with this compound at POCl ₃With heat in the pyridine, can produce and comprise Δ ^9,11The compound 167 and the Δ thereof of two keys ^11,12Isomer.Adopt mCPBA that compound 167 is carried out epoxidation, use LiAlH again ₄Reduce, can obtain comprising the compound 169 of C9 hydroxyl functional base.This hydroxyl is protected, removed de ketal protecting group again, and carry out Wittig reaction, can obtain olefin product 171.Adopt standard method described herein compound 171 can be changed into lactonaphthol 176.

Second example relates to preparation derivative 186, and this compound comprises C15 ketone and side chain hemiacetal and C1 hydroxyl functional base.Can prepare compound 186 through multistep process by commercially available raw material 177, shown in reaction scheme 48.The first step comprises the benzole soln protection compound 178 of employing such as ethylene glycol, pTsOH.Subsequently, for example adopt that benzylalcohol and potassium hydroxide carry out Michael reaction, obtain C1 benzyloxy derivatives 179.Ketone 179 is carried out LS-Selectride  reduction, again the alcohol that forms is protected with benzyloxy derivatives, obtain compound 180.Then, adopt reaction scheme 47 described methods to reach the method that in other previous examples, describes in detail and compound 180 can be changed into lactonaphthol 186.

Reaction scheme 48

Therefore, method described here can be used for being created on the carbon of cyclopentanoperhydro-phenanthrene structure and has functional group and have C15 ketone functional group and the compound of side chain hemiacetal.

The present invention also provides a kind of and has had oxide group at C6 and C7 position, has in the C17 position to contain pyrans-or the steroide of delta-lactone side chain.In one embodiment, this steroide has structural formula 11, comprises independent enantiomorph or its geometrical isomer, also comprises its solvate or pharmacy acceptable salt.Structural formula 11 is defined as follows:

Compound with following formula

Comprise its pharmacy acceptable salt and solvate, wherein:

G is-C (=O)-,-CH (OR ¹)-,-C (R ⁴) (OR ¹)-or-C (OR ¹) (OR ¹)-;

X represents fluorine, chlorine, bromine and iodine.

The preparation of structural formula 12 compounds can be adopted many local methods that propose herein.For example, compound 196 (reaction scheme 49) and 207 (reaction schemes 50) can be synthetic in the rapid method of multistep by compound 30 and 55.Being used for C17 siloxy-with compound 30, to change into the method for describing in detail in the method for alkene 190 and the previous examples similar, as be used for compound 190 is changed into the method for compound 196.It also is like this respectively compound 55 being changed into compound 200 and changing into compound 207 from compound 200.

Reaction scheme 49

Reaction scheme 50

In the as above reaction described in the reaction scheme 49 and 50 only is how method that the present invention discussed is applied to produce and contains 6, two examples of the compound of 7-titanium dioxide form and hemiacetal or delta-lactone side chain.Therefore, aforesaid method can be used for producing the compound with functional groups such as C2, C4, C8.

The present invention also provides the epoxide of steroide.In one embodiment, the steroide epoxide has structural formula 12, comprises independent enantiomorph or its geometrical isomer, also comprises its solvate or pharmacy acceptable salt.Structural formula 12 is defined as follows:

Compound with following formula

Comprise its pharmacy acceptable salt and solvate, wherein:

X represents fluorine, chlorine, bromine and iodine;

Example as described above, adopt methods described herein can structural formula 12 contain 3, functional group is introduced in each position in the steroide ring structure of 4-epoxide group.For example, shown in reaction scheme 51, through Δ ^9,11The epoxidation of two keys, Sauerstoffatom can be placed in C9 and/or C11 position.

Therefore, after the LS-Selectride reduction, adopting as previously mentioned, reagent can obtain olefin(e) compound 208 (reaction scheme 51) to carrying out far-end (remote) oxidation such as compound 10 at interior compound.Adopt standard method can be converted into Δ ^9,11Alkene, Δ ^3,4And Δ ^9,11Two keys react simultaneously, obtain the required epoxide between C3-C4 and C9-C11.Then, use the CH of PDC ₂Cl ₂Solution oxide C3 hydroxyl obtains required unsaturated A-ring (and, oxirane ring is carried out the optional open loop, obtain 3,6,7, the polyhydroxylated steroide 215 of 9-).

Reaction scheme 51

Adopt similar as previously mentioned reaction also can introduce alkyl in the C16 position.In following example (reaction scheme 52), introduce methyl in this position by making D-cyclenes alkoxide and methyl-iodide condensation.Method is similar shown in this method and the reaction scheme 37.Shown in reaction scheme 52, alkylating epoxide 218 can place under the epoxide open loop condition, obtains 3,4,6,7-tetrahydroxy steroide 220.

Reaction scheme 52

Gone through in front and had 6 α, the compound of 7 beta-hydroxy forms.In addition, as following will the discussion, also can produce and comprise the stereomeric compound of other C6 and C7 position.For example, adopt the pyridine solution of pTsCl that compound 221 (preparing according to reaction scheme 73) is carried out selectivity tosylation reaction, handle with salt of wormwood again, obtain epoxidised compound 223.Subsequently, adopt aqueous acids to carry out ring-opening reaction, can obtain having 6 β, the stereomeric compound of 7 α is shown in reaction scheme 53.

Reaction scheme 53

Have 6 α, the stereomeric compound of 7 α can be from commercially available feedstock production, shown in reaction scheme 54.Rhabdasterol acetate can be used RuCl ₃CH with tBuOOH ₂Cl ₂Solution oxide obtains containing the compound 229 of ketenes.Protecting group with 3 is exchanged for the tBDMS derivative, again with lithium/ammonia reduction, with (MeO) ₂PCl catches the enolate negatively charged ion, obtains enol phosphate 231.Lithium-ammonia reduction for the second time obtains Δ ^6,7Two keys are used OsO with it ₄Oxidation obtains containing 3 β, 6 α, the compound 233 of 7 α-trihydroxy-form.

Reaction scheme 54

The term of used appointment carbon atom number range " organic residue " is meant the stable alignment of atom among the present invention, described atom is by at least 1 carbon atom and is no more than the maximum carbonatoms of the scope that proposes that the non-carbon atom that is no more than about 30 carbon atoms and any amount is usually formed.

C ₁-C ₃₀Organic residue can be saturated or unsaturated alkyl.Saturated hydrocarbyl of the present invention is defined as any group of only being made up of carbon and hydrogen, wherein, only adopts singly-bound to connect carbon atom.Therefore, the carbon of any stable alignment that has 1 carbon atom at least and hydrogen atom are all in the range of definition of saturated hydrocarbyl of the present invention.Below discussion is used to censure some specific nomenclature of particular carbon atomic arrangement.

Carbon atom can form alkyl, and the carbon atom that promptly can be side chain or non-side chain (straight chain) does not have loop chain.Methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, isobutyl-, sec-butyl and the tertiary butyl are the alkyl (being referred to as low alkyl group usually) with 1-4 carbon atom, and they also are the examples of alkyl of the present invention.Carbon atom can form cycloalkyl, i.e. the circular permutation of carbon atom, and cyclopropyl, cyclobutyl, cyclopentyl are the cycloalkyl that the present invention has 3-5 carbon atom.Other group that the present invention defines in " cycloalkyl " scope is below with the multi-ring alkyl that defines.

Multi-ring alkyl is a kind of arrangement mode of carbon atom, and wherein, at least one carbon atom is the part of at least two diacritic rings that separate.Multi-ring alkyl can comprise two bridged bonds between carbon atom, and wherein, representative example is dicyclo [1.1.0] butyl, dicyclo [3.2.1] octyl group, dicyclo [5.2.0] nonyl, three ring [2.2.1.0 ¹] heptyl, norborneol alkyl (norbornyl) and pinane base (pinanyl).Multi-ring alkyl can comprise one or more fused rings systems, and wherein, representative example is decahydro naphthyl (coming from perhydronaphthalene) and perhydro anthryl.Multi-ring alkyl can comprise and is spirally connected, and a wherein single atom is the unique total member of two rings.Representative example is spiral shell [3.4] octyl group, spiral shell [3.3] heptyl and spiral shell [4.5] decyl.

In addition, saturated hydrocarbyl can be formed by two or more above-mentioned group arbitrary combination, i.e. the arbitrary combination of alkyl and cycloalkyl.Therefore, R ⁴Or R ⁵Group can be and has the substituent alkyl of cycloalkyl (as cyclohexyl) (as methyl), thereby, R ⁴Or R ⁵Be cyclohexyl methyl.In another example, R ⁴Or R ⁵Group can be have two alkyl substituents (as methyl and ethyl substituting group) cycloalkyl (as the ring octyl group), thereby, R ⁴Or R ⁵Be methylethyl ring octyl group.As last example, R ⁴Or R ⁵Can be the cycloalkyl that has alkyl substituent, wherein, alkyl substituent is replaced by the multi-ring alkyl substituting group.

As mentioned above, R ⁴Or R ⁵Can be unsaturated alkyl.This R ⁴Or R ⁵Group definition is to have as preceding the carbon that saturated hydrocarbyl proposed is arranged, and supplementary features are that at least one key between any two carbon atoms is not a singly-bound.Alkyl with two keys is called as alkenyl, and the alkyl that has more than two keys is called as alkapolyenyl (alkapolyenyl), and its specific examples is alkadienyl (2 two keys) and alkatriene base (3 two keys).Alkyl with one three key is called as alkynyl group, and the alkyl that has more than one three key is called as chain polyyne base, and its specific examples is alkadiyne base (2 three keys) and alkatriyne base (3 three keys).

Similarly, cyclic hydrocarbon radical can have one or more pairs of keys or three key, and they are also included within the unsaturated alkyl of the present invention.Cycloalkenyl group and cycloalkynyl radical are the common names that has the single carbon ring group of two keys and one three key in the ring respectively.Cycloalkadienyl is for having the cyclic hydrocarbon radical of two two keys in ring structure.Two keys can be exocyclic double bond, and for example, available ring carbon atom has=CH ₂Group (that is methylene radical) or the homologue that is attached thereto of high-grade more.

The degree of unsaturation of ring can be to aromaricity, and they still are included in the unsaturated alkyl scope.Therefore, aryl such as phenyl and naphthyl are included within this alkyl scope.Above-mentioned any array mode is also included within the unsaturated alkyl scope, R ⁴Or R ⁵Comprise aralkyl (R ⁴Or R ⁵For having the alkyl of an aryl substituent at least, as benzyl) and alkaryl (R ⁴Or R ⁵For having the aryl of an alkyl substituent at least, as tolyl).The preferred embodiment of the organic residue of the present invention is C ₆Aryl.

R ⁴Or R ⁵Comprise and contain heteroatomic organic residue.Heteroatoms of the present invention is any atom outside de-carbon and the hydrogen atom.Preferred heteroatoms is naturally occurring atom (except that carbon atom and a hydrogen atom).Another kind of preferred heteroatoms is non-metallic atom (except that carbon atom and a hydrogen atom).Another kind of preferred heteroatoms is made up of boron, nitrogen, oxygen, silicon, phosphorus, sulphur, selenium and halogen (being fluorine, chlorine, bromine and iodine, preferred fluorine and chlorine).Another kind of preferred heteroatoms is made up of nitrogen, oxygen, sulphur and halogen.Another kind of preferred heteroatoms is made up of nitrogen, oxygen and sulphur.Oxygen is preferred heteroatoms.Nitrogen is preferred heteroatoms.

For example, R ⁴Or R ⁵Can be the alkyl as preceding definition, it has one at least and contains at least one heteroatomic substituting group.At this section, R ⁴To be used to represent R ⁴And R ⁵That is to say R ⁴Can be the alkyl as preceding definition, wherein, at least one hydrogen atom is replaced by a heteroatoms.For example, if heteroatoms is an oxygen, then substituting group can be carbonyl, and promptly two hydrogen on a carbon atom are replaced by oxo, forms ketone or aldehyde group.In addition, a hydrogen atom can be replaced by a former following radicals form that gives of oxygen, described form is hydroxyl, alkoxyl group, aryloxy, aralkoxy, aryloxy alkyl (wherein, alkoxyl group, aryloxy, aralkoxy, aryloxy alkyl can be referred to as-oxyl), heteroaryloxy, OC (O) R ⁴, ketal, acetal, hemiketal, hemiacetal, epoxidation and-OSO ₃M.Heteroatoms can be halogen atom.Heteroatoms can be nitrogen, and wherein nitrogen becomes the part of following radicals: amino (NH ₂,-NHR ₄, N (R ⁴) ₂), alkyl amido, aryl amido, aralkyl amido, alkaryl amido, nitro ,-N (R ⁴) SO ₃M or aminocarboxyl amide group.Heteroatoms can be sulphur, wherein, sulphur form mercaptan, thiocarbonyl ,-SO ₃The part of M, alkylsulfonyl, sulfamyl or sulfonyl hydrazino.Heteroatoms can be carbon containing substituting group such as formyl radical, cyano group ,-C (O) OR ⁴,-C (O) OM ,-C (O) R ⁴,-C (O) N (R ⁴) ₂, carbamate, carbohydrazide and carbon hydroxamic acid a part.

Contain in the hetero atom substituents example above-mentioned, M represents proton or metal ion.Preferred metal ion and counter ion are combined to form the physiological salt that tolerates.Preferable alloy ionic metal be can obtain by it and basic metal [for example: lithium (Li), sodium (Na), potassium (K), rubidium (Rb) and caesium (Cs)], alkaline-earth metal (for example, magnesium (Mg), calcium (Ca) and strontium (Sr)), manganese (Mn), iron (Fe), zinc (Zn) or silver (Ag) comprised.Basic metal or alkaline-earth metal are the preferred L groups.Sodium, potassium, magnesium and calcium are the preferred L groups.Sodium and potassium are the preferred L groups.

Another kind of organic residue of the present invention is the alkyl as preceding definition, and wherein, at least one carbon atom is replaced by at least one heteroatoms.The example of this organic residue is Heterocyclylalkyl (having a carbon atom at least by the displaced cycloalkyl of at least one heteroatoms), heterocycloalkenyl, heteroaryl, heteroaryl oxygen base, heteroaralkyl, impure aromatic ene base etc.In general, the organic residue of this class can be called as assorted alkyl.The example of another kind of this organic residue has a heteroatoms, the group that this heteroatoms bridging (a) links to each other with organic residue and (b) other parts of organic residue.The example comprises alkoxyl group, aryloxy, alkoxy aryl and alkyl-aryloxy, and they are referred to as-oxyl in this article.Therefore, R of the present invention ⁴An example of group is-OR ⁴Another example is-NHR ⁴

The example of inferior Heterocyclylalkyl is pyrrolidinylidene, piperidylidene, inferior tetrahydrofuran base, inferior dihydro pyranyl and inferior THP trtrahydropyranyl.The example of Heterocyclylalkyl is by following substances deutero-group: tetramethyleneimine, imidazolidine, oxazolidine, pyrazolidine, piperidines, piperazine and morpholine.The substituent example of heterocycloalkenyl is by deutero-group behind 2-and 3-pyrroline, oxazoline, 2-and 4-tetrahydroglyoxaline and 2-and hydrogen of 3-pyrazoline removal.

Organic residue can have about at the most 30 carbon atoms, and preferred organic residue of the present invention has less than 30 carbon atoms for example about at the most 25 carbon atoms, preferably about at the most 20 carbon atoms.Organic residue can have about at the most 15 carbon atoms, or about at the most 12 or 10 carbon atoms.Preferred organic residue has about at the most 8 or 6 carbon atoms.

Below be the R of illustrative ⁴And R ⁵Organic residue, wherein, R ⁴Or R ⁵Be connected on the steroid nucleus by carbon atom: alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group, aryl, heterocyclic radical, alkyl-carbonyl, alkenyl carbonyl, alkynyl group carbonyl, naphthene base carbonyl, cycloalkenyl carbonyl, aryl carbonyl, heterocyclic radical carbonyl, alkoxy carbonyl, alkenyloxy carbonyl, chain oxy-acetylene carbonyl, cyclo alkoxy carbonyl, ring allyloxycarbonyl, aryloxy carbonyl, heterocyclyloxy base carbonyl, carboxylic acid, cyano group and formyl radical.

Below be the R of illustrative ⁴And R ⁵Organic residue, wherein, R ⁴Or R ⁵Be connected on the steroid nucleus by Sauerstoffatom: hydroxyl, oxo, alkoxyl group, alkenyloxy, chain oxy-acetylene, cycloalkyloxy, cyclenes oxygen base, aryloxy, alkyl-carbonyl oxygen base, alkenyl carbonyl oxygen base, alkynyl group ketonic oxygen base, naphthene base carbonyl oxygen base, cycloalkenyl carbonyl oxygen base, aryl carbonyl oxygen base and heterocyclyloxy base.

R ⁴And R ⁵Can comprise a nitrogen-atoms, by this nitrogen-atoms, R ⁴Or R ⁵Organic residue is connected on the steroid nucleus.The example is a nitro, formula NL ²L ³Organic residue, L wherein ²And L ³Be independently of one another: hydrogen, alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group, aryl, formyl radical, heterocyclic radical, alkyl-carbonyl, alkenyl carbonyl, alkynyl group carbonyl, naphthene base carbonyl, cycloalkenyl carbonyl, aryl carbonyl and heterocyclic radical carbonyl; thereby, L ²And L ³Can be alkylidene group or alkylene group together, with the combination of the nitrogen-atoms that links to each other with it, forming 3-unit does not have saturated or unsaturated ring to 8-.

Below be the R of illustrative ⁴And R ⁵Organic residue, wherein, R ⁴Or R ⁵Be connected on the steroid nucleus by sulphur atom: the group of alkylthio, alkenyl thio, alkynyl group sulfenyl, cycloalkylthio, cyclenes sulfenyl, arylthio, heterocycle sulfenyl, alkyl-carbonyl sulfenyl, alkenyl carbonyl sulfenyl, alkynyl group carbonyl sulfenyl, naphthene base carbonyl sulfenyl, cycloalkenyl carbonyl sulfenyl, aryl carbonyl sulfenyl, heterocyclic radical carbonyl sulfenyl and following formula :-S (O) _nH ,-S (O) _nL ⁴,-S (O) _mOH ,-S (O) _mOL ⁴, OS (O) _mOL ⁴With-O (S) _mOH, L ⁴Be selected from alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group, aryl and heterocyclic radical.

At above R ⁴And R ⁵In organic residue, alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group and aryl (are referred to as R ⁴Or R ⁵Alkyl) can be by all halogenations or partially halogenated, and/or with five L at the most ⁵Group replaces.Similarly, heterocyclic radical, heterocyclyloxy base, heterocyclic radical carbonyl, heterocyclyloxy base carbonyl, heterocyclic radical ketonic oxygen base (are referred to as R ⁴Heterocyclic radical) can be by all halogenations or partially halogenated, and/or with five L at the most ⁵Group replaces.

L ⁵Group comprises carbon, oxygen, nitrogen or sulphur atom, by these atoms, and they and R ⁴Or R ⁵The carbon atom of alkyl links to each other, perhaps with R ⁴Or R ⁵The carbon of heterocyclic radical or nitrogen-atoms link to each other.

Below be the L of illustrative ⁵Group, wherein, L ⁵Carbon atom and R ⁴Alkyl or heterocyclic radical link to each other: alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group, aryl, alkyl-carbonyl, alkenyl carbonyl, alkynyl group carbonyl, naphthene base carbonyl, cycloalkenyl carbonyl, aryl carbonyl, alkoxy carbonyl, alkenyloxy carbonyl, chain oxy-acetylene carbonyl, cyclo alkoxy carbonyl, ring allyloxycarbonyl and aryloxycarbonyl.

Below be the L of illustrative ⁵Group, wherein, L ⁵Sauerstoffatom and R ⁴Alkyl or heterocyclic radical link to each other: hydroxyl, oxo, alkoxyl group, alkenyloxy, chain oxy-acetylene, cycloalkyloxy, cyclenes oxygen base, aryloxy, alkyl-carbonyl oxygen base, alkenyl carbonyl oxygen base, alkynyl group ketonic oxygen base, naphthene base carbonyl oxygen base, cycloalkenyl carbonyl oxygen base and aryl carbonyl oxygen base.

L ⁵Group can comprise nitrogen-atoms, by this nitrogen-atoms, L ⁵Group and R ⁴Or R ⁵Alkyl or heterocyclic radical link to each other.The example comprises nitro and formula-NL ⁶L ⁷Nitrogen-containing group, L wherein ⁶And L ⁷Be hydrogen, alkyl, alkenyl, alkynyl group, cycloalkyl, aryl, formyl radical, alkyl-carbonyl, alkenyl carbonyl, alkynyl group carbonyl, naphthene base carbonyl, cycloalkenyl carbonyl and aryl carbonyl independently, L ⁶And L ⁷Can be alkylidene group or alkylene group together, thereby be combined to form the saturated or unsaturated ring of 3-to 8-unit with the nitrogen-atoms that is attached thereto.

Below be the L of illustrative ⁵Group, wherein, L ⁵Sulphur atom and R ⁴Or R ⁵Alkyl or heterocyclic radical link to each other: the group of alkylthio, alkenyl thio, alkynyl group sulfenyl, cycloalkylthio, cyclenes sulfenyl, arylthio, alkyl-carbonyl sulfenyl, alkenyl carbonyl sulfenyl, alkynyl group carbonyl sulfenyl, naphthene base carbonyl sulfenyl, cycloalkenyl carbonyl sulfenyl, aryl carbonyl sulfenyl and following formula :-S (O) _nL ⁸,-S (O) _mOH ,-S (O) _mOl ⁸, OS (O) _mOL ⁸With-O (S) _mOH, L ⁸Be selected from alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group, aryl and heterocyclic radical.

R in illustrative ⁴And R ⁵In organic residue, form L ⁵Alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group and the aryl of part (are referred to as L ⁵Alkyl) all halogenations or partially halogenated, and/or with three L at the most ⁹Group replaces.Similarly, heterocyclic radical, heterocyclyloxy base, heterocyclic radical carbonyl, heterocyclyloxy base carbonyl, heterocyclic radical ketonic oxygen base (are referred to as L ⁵Heterocyclic radical) all halogenations or partially halogenated, and/or with three L at the most ⁹Group replaces.

L ⁹Group comprises carbon, oxygen, nitrogen or sulphur atom, by these atoms, and they and L ⁵Alkyl or L ⁵Heterocyclic radical links to each other.

Below be the L of illustrative ⁹Group, wherein, L ⁹Carbon atom and L ⁵Alkyl or heterocyclic radical link to each other: alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group, aryl, heterocyclic radical, alkyl-carbonyl, alkenyl carbonyl, alkynyl group carbonyl, naphthene base carbonyl, cycloalkenyl carbonyl, aryl carbonyl, heterocyclic radical carbonyl, alkoxy carbonyl, alkenyloxy carbonyl, chain oxy-acetylene carbonyl, cyclo alkoxy carbonyl, ring allyloxycarbonyl, aryloxycarbonyl and heterocyclyloxy base carbonyl.

Below be the L of illustrative ⁹Group, wherein, L ⁹Sauerstoffatom and L ⁵Alkyl or heterocyclic radical link to each other: hydroxyl, oxo, alkoxyl group, alkenyloxy, chain oxy-acetylene, cycloalkyloxy, cyclenes oxygen base, aryloxy, heterocyclyloxy base, alkyl-carbonyl oxygen base, alkenyl carbonyl oxygen base, alkynyl group ketonic oxygen base, naphthene base carbonyl oxygen base, cycloalkenyl carbonyl oxygen base, aryl carbonyl oxygen base and heterocyclic radical ketonic oxygen base.

L ⁹Group can comprise nitrogen-atoms, by this nitrogen-atoms, L ⁹Group and L ⁵Alkyl or heterocyclic radical link to each other.This nitrogenous L ⁹Examples of groups comprises nitro and formula-NL ¹⁰L ¹¹Group, L wherein ¹⁰And L ¹¹Be hydrogen, alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group, aryl, heterocyclic radical, formyl radical, alkyl-carbonyl, alkenyl carbonyl, alkynyl group carbonyl, naphthene base carbonyl, cycloalkenyl carbonyl, aryl carbonyl and heterocyclic radical carbonyl independently, L ¹⁰And L ¹¹Can be alkylidene group or alkylene group together, thereby be combined to form the saturated or unsaturated ring of 3-to 8-unit with the nitrogen-atoms that is attached thereto.

Below be the L of illustrative ⁹Group, wherein, L ⁹Sulphur atom and L ⁵Alkyl or heterocyclic radical link to each other: the group of alkylthio, alkenyl thio, alkynyl group sulfenyl, cycloalkylthio, cyclenes sulfenyl, arylthio, heterocyclic radical sulfenyl, alkyl-carbonyl sulfenyl, alkenyl carbonyl sulfenyl, alkynyl group carbonyl sulfenyl, naphthene base carbonyl sulfenyl, cycloalkenyl carbonyl sulfenyl, aryl carbonyl sulfenyl, heterocyclic radical carbonyl sulfenyl and following formula :-S (O) _nL ¹²,-S (O) _mOH ,-S (O) _mOL ¹², OS (O) _mOL ¹²With-O (S) _mOH, L ¹²Be selected from alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group, aryl and heterocyclic radical.

At R ⁴And R ⁵In the example of organic residue, form L ⁹Alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group and the aryl of part (are referred to as L ⁹Alkyl) all halogenations or partially halogenated, and/or with three L at the most ¹³Group replaces.Similarly, heterocyclic radical, heterocyclyloxy base, heterocyclic radical carbonyl, heterocyclyloxy base carbonyl, heterocyclic radical ketonic oxygen base (are referred to as L ⁹Heterocyclic radical) all halogenations or partially halogenated, and/or with three L at the most ¹³Group replaces.

L ¹³Group comprises carbon, oxygen, nitrogen or sulphur atom, by these atoms, L ¹³With L ⁹Alkyl or L ⁹Heterocyclic radical links to each other.

Below be the L of illustrative ¹³Group, wherein, L ¹³Carbon atom and L ⁹Alkyl or heterocyclic radical link to each other: alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group, aryl, heterocyclic radical, alkyl-carbonyl, alkenyl carbonyl, alkynyl group carbonyl, naphthene base carbonyl, cycloalkenyl carbonyl, aryl carbonyl, heterocyclic radical carbonyl, alkoxy carbonyl, alkenyloxy carbonyl, chain oxy-acetylene carbonyl, cyclo alkoxy carbonyl, ring allyloxycarbonyl, aryloxycarbonyl and heterocyclyloxy base carbonyl.

Below be the L of illustrative ¹³Group, wherein, L ¹³Sauerstoffatom and L ⁹Alkyl or heterocyclic radical link to each other: hydroxyl, oxo, alkoxyl group, alkenyloxy, chain oxy-acetylene, cycloalkyloxy, cyclenes oxygen base, aryloxy, heterocyclyloxy base, alkyl-carbonyl oxygen base, alkenyl carbonyl oxygen base, alkynyl group ketonic oxygen base, naphthene base carbonyl oxygen base, cycloalkenyl carbonyl oxygen base, aryl carbonyl oxygen base and heterocyclic radical ketonic oxygen base.

L ¹³Group can comprise nitrogen-atoms, by this nitrogen-atoms, L ¹³Group and L ⁹Alkyl or heterocyclic radical link to each other.This nitrogenous L ¹³Examples of groups comprises nitro and formula-NL ¹⁴L ¹⁵Group, L wherein ¹⁴And L ¹⁵Be hydrogen, alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group, aryl, heterocyclic radical, formyl radical, alkyl-carbonyl, alkenyl carbonyl, alkynyl group carbonyl, naphthene base carbonyl, cycloalkenyl carbonyl, aryl carbonyl and heterocyclic radical carbonyl independently, L ¹⁴And L ¹⁵Can be alkylidene group or alkylene group together, thereby be combined to form the saturated or unsaturated ring of 3-to 8-unit with the nitrogen-atoms that is attached thereto.

Below be the L of illustrative ¹³Group, wherein, L ¹³Sulphur atom and L ⁹Alkyl or heterocyclic radical link to each other: the group of alkylthio, alkenyl thio, alkynyl group sulfenyl, cycloalkylthio, cyclenes sulfenyl, arylthio, heterocyclic radical sulfenyl, alkyl-carbonyl sulfenyl, alkenyl carbonyl sulfenyl, alkynyl group carbonyl sulfenyl, naphthene base carbonyl sulfenyl, cycloalkenyl carbonyl sulfenyl, aryl carbonyl sulfenyl, heterocyclic radical carbonyl sulfenyl and following formula :-S (O) _nL ¹⁴,-S (O) _mOH ,-S (O) _mOL ¹⁴, OS (O) _mOL ¹⁴With-O (S) _mOH, L ¹⁴Be selected from alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group, aryl and heterocyclic radical.

In above-mentioned group, m is l or 2 independently, and n is 0,1 or 2 independently.

Some R ⁴And R ⁵Substituting group can comprise unsymmetrical carbon.Thereby, comprise this substituent compound and have enantiomorph and diastereomeric form and racemic mixture thereof.All these forms is all within the scope of the invention.Racemic modification or racemic mixture also do not mean that to be 50: 50 stereoisomer mixture.

Relevant to specifications R ⁴Or R ⁵The illustrative of organic residue is described, and except as otherwise noted, following term has following implication.

Alkyl, alkenyl and alkynyl group be meant do not have unsaturated link(age) respectively, have at least one two key or have at least one three key, comprise 1-30 the carbon atom straight or branched alkyl of (alkynyl group comprises 2 carbon atoms at least).

Cycloalkyl and cycloalkenyl group are meant the cyclic hydrocarbon radical that comprises 3-8 carbon atom, and wherein, cycloalkyl is saturated, and cycloalkenyl group has the two keys at least one ring structure.Suitable cycloalkyl comprises cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, suberyl and ring octyl group.

Aryl is meant and comprises an aromatic group with ring of conjugated pi electron system at least, comprises isocyclic aryl, heterocyclic aryl and dibenzyl.

Isocyclic aryl is meant that the annular atoms of aromatic ring is the aryl of carbon atom.Isocyclic aryl comprises phenyl, naphthyl and indenyl.

Heterocyclic aryl is meant list or the bicyclic system that comprises about 12 carbon atoms of about 5-, wherein, each monocycle has about 4 heteroatomss of 0-, and each dicyclo has about 5 heteroatomss of about 0-, heteroatoms is selected from N, O and S, and condition is that described heteroatoms is not ortho position oxygen and/or sulphur atom.The example of this list and bicyclic system includes but not limited to: cumarone, thionaphthene, indoles, benzopyrazoles, tonka bean camphor, isoquinoline 99.9, pyrroles, thiophene, furans, thiazole, imidazoles, pyrazoles, thiazole, quinoline, pyrimidine, pyridine, pyridone, pyrazine, pyridazine, isothiazole, isoxazole and tetrazolium.

Dibenzyl is meant foregoing by the phenyl of isocyclic aryl or heterocyclic aryl replacement, and the position of substitution can be in ortho position, a position or the contraposition of phenyl ring tie point.

Heterocyclic radical is meant stable 5-to 7-unit's monocycle or the two heterocyclic systems of dicyclo or 7-to 10-unit, arbitrary ring can be saturated or undersaturated, it is made up of carbon atom and 1-3 heteroatoms, described heteroatoms is selected from N, O and S, wherein, nitrogen and sulphur atom can be optionally oxidized, and nitrogen heteroatom can be optionally by quaternized, and comprise any above-mentioned heterocycle and phenyl ring condensed dicyclo.Heterocycle can be connected with steroid nucleus by any heteroatoms on the heterocycle or carbon atom, as long as form stable structure.The example of this heterocyclic group comprises: piperidyl, piperazinyl, 2-oxo piperazinyl, 2-oxo-piperidine base, 2-oxo-pyrrolidine base, 2-oxo azatropylidene base, the azatropylidene base, pyrryl, the 4-piperidone base, pyrrolidyl, pyrazolyl, pyrazolidyl, imidazolyl, imidazolinyl, imidazolidyl, pyridyl, pyrazinyl, pyrimidyl, pyridazinyl oxazolyl Evil oxazolidinyl Yi Evil oxazolyl isoxazole alkyl, morpholinyl, thiazolyl, thiazolidyl, isothiazolyl, quinuclidinyl, the isothiazole alkyl, indyl, quinolyl, isoquinolyl, benzimidazolyl-, thiadiazolyl group, benzopyranyl, benzothiazolyl benzoxazolyl, furyl, tetrahydrofuran base, THP trtrahydropyranyl, thienyl, benzothienyl, the parathiazan base, the parathiazan sulfoxide, parathiazan Feng is with oxadiazole base base.Morpholino (morpholino) is identical with morpholinyl (morpholinyl).

Heterocyclyloxy base and heterocyclic radical carbonyl are meant respectively by Sauerstoffatom or carbonyl and one or more steroid nucleus, R ⁴Alkyl, L ⁵Alkyl or L ⁹The heterocyclic radical that one of alkyl links to each other.

Heterocyclyloxy base carbonyl is meant by carbonyl and one or more steroid nucleus, R ⁴Alkyl, L ⁵Alkyl or L ⁹The heterocyclyloxy base that one of alkyl links to each other.

Heterocyclic radical ketonic oxygen base is meant by Sauerstoffatom and one or more steroid nucleus, R ⁴Alkyl, L ⁵Alkyl or L ⁹The heterocyclic radical carbonyl that one of alkyl links to each other.

Alkyl-carbonyl, alkenyl carbonyl, alkynyl group carbonyl, naphthene base carbonyl, cycloalkenyl carbonyl and aryl carbonyl are meant following group: wherein carbonyl (C=O) provides carbon atom, by described carbon atom, and described group and steroid nucleus, R ⁴Alkyl, L ⁵Alkyl or L ⁹One of alkyl links to each other, and alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group or aryl also link to each other with carbonyl respectively.

Alkoxy carbonyl, alkenyloxy carbonyl, chain oxy-acetylene carbonyl, cyclo alkoxy carbonyl, ring allyloxycarbonyl and aryloxycarbonyl are meant following group: wherein carbonyl (C=O) provides carbon atom, by described carbon atom, and described group and steroid nucleus, R ⁴Alkyl, L ⁵Alkyl or L ⁹One of alkyl links to each other, and alkoxyl group, alkenyloxy, chain oxy-acetylene, cycloalkyloxy, cyclenes oxygen base or aryloxy also divide other to link to each other with carbonyl.

Alkoxyl group, alkenyloxy, chain oxy-acetylene, cycloalkyloxy, cyclenes oxygen base and aryloxy are meant following group: wherein, oxygen links to each other with alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group or aryl respectively, and oxygen also with steroid nucleus, R ⁴Alkyl, L ⁵Alkyl or L ⁹One of alkyl links to each other.

Alkyl-carbonyl oxygen base, alkenyl carbonyl oxygen base, alkynyl group ketonic oxygen base, naphthene base carbonyl oxygen base, cycloalkenyl carbonyl oxygen base and aryl carbonyl oxygen base are meant following group: wherein, oxygen respectively with alkyl-carbonyl, alkenyl carbonyl, alkynyl group carbonyl, naphthene base carbonyl, cycloalkenyl carbonyl or aryl carbonyl, link to each other and oxygen also with steroid nucleus, R ⁴Alkyl, L ⁵Alkyl or L ⁹One of alkyl links to each other.

Alkylthio, alkenyl thio, alkynyl group sulfenyl, cycloalkyl sulfenyl, cycloalkenyl group sulfenyl and artyl sulfo are meant following group: wherein, sulphur content does not link to each other with alkyl, alkenyl, alkynyl group, cycloalkyl, cycloalkenyl group or aryl, and sulphur also with steroid nucleus, R ⁴Alkyl, L ⁵Alkyl or L ⁹One of alkyl links to each other.

Alkyl-carbonyl sulfenyl, alkenyl carbonyl sulfenyl, alkynyl group carbonyl sulfenyl, naphthene base carbonyl sulfenyl, cycloalkenyl carbonyl sulfenyl and aryl carbonyl sulfenyl are meant following group: wherein, sulphur content does not link to each other with alkyl-carbonyl, alkenyl carbonyl, alkynyl group carbonyl, naphthene base carbonyl, cycloalkenyl carbonyl or aryl carbonyl, and sulphur also with steroid nucleus, R ⁴Alkyl, L ⁵Alkyl or L ⁹One of alkyl links to each other.

Alkylidene group is meant the straight chain bridge with 1-5 carbon atom, and it can be replaced by 1-3 low alkyl group or all or part of halogenated low alkyl group.

Alkylene group is meant the straight chain bridge that has 2-5 carbon atom and have 1 or 2 two keys, and it can be replaced by 1-3 low alkyl group or all or part of halogenated low alkyl group.

Inferior alkynyl group is meant the straight chain bridge that has 2-5 carbon atom and have 1 or 2 three key, and it can be replaced by 1-3 low alkyl group or all or part of halogenated low alkyl group.

Low alkyl group is meant the C1-C5 alkyl, as methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, the tertiary butyl, sec-butyl, isobutyl-, n-pentyl, isopentyl etc.

Halogen is meant fluorine, chlorine, bromine and iodine, and the halo group is meant the carbon atom that is connected with at least 1 halogen atom.

Formyl radical is meant-C (=O) H; Hydroxyl is meant-OH; Oxo is meant the Sauerstoffatom that constitutes carbonyl moiety.

Pharmacy acceptable salt comprises acid salt and base addition salt.

Acid salt is meant the salt that is formed by steroide of the present invention and mineral acid and/or organic acid, described mineral acid for example is hydrochloric acid, Hydrogen bromide, sulfuric acid, nitric acid, phosphoric acid etc., and described organic acid for example is acetate, propionic acid, oxyacetic acid, pyruvic acid, oxalic acid, toxilic acid, propanedioic acid, succsinic acid, fumaric acid, tartrate, citric acid, phenylformic acid, styracin, amygdalic acid, methylsulfonic acid, ethyl sulfonic acid, tosic acid, Whitfield's ointment etc.

Base addition salt comprises the salt that is formed by steroide of the present invention and mineral alkali, for example sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminium salt etc.Suitable salt comprises the ammonium that is obtained by pharmaceutically acceptable organic nontoxic alkali, potassium, sodium, calcium and magnesium salts, the salt that comprises following substances: primary, the second month in a season and tertiary amine, the amine that replaces comprises the amine of natural replacement, cyclammonium and deacidite are as Isopropylamine, Trimethylamine 99, diethylamine, triethylamine, tripropyl amine, thanomin, the 2-dimethylaminoethanol, the 2-DEAE diethylaminoethanol, trimethamine, dicyclohexyl amine, Methionin, arginine, Histidine, caffeine, PROCAINE HCL, PHARMA GRADE, Hydrabamine Peniccilin G (hydrabamine), choline, trimethyl-glycine, quadrol, glycosamine, methylglucosamine, Theobromine, purine, piperazine, piperidines, N-ethylpiperidine etc.

In another embodiment, the invention provides a kind of composition, it comprises aforesaid 6,7-titanium dioxide steroide and mixing with it or otherwise one or more inert supports of bonded and the optional member when needing.A kind of pharmaceutical composition, it comprises a kind of compound and a kind of pharmaceutically acceptable carrier or thinner, and this compound has following formula

Comprise its pharmacy acceptable salt and solvate, wherein:

C5, C6, C7, C8, C9, C10, C13 and C14 independently of one another by-X ,-R ⁴With-OR ¹Replace;

C1, C2, C3, C4, C11, C12, C15, C16 and C17 following by being selected from independently of one another (a) or substituting group (b) replace:

(a) represent one of following formula :=O ,=C (R ⁴) (R ⁴) ,-C (R ⁴) (R ⁴) (C (R ⁴) (R ⁴)) _n-and-(O (C (R ⁴) (R ⁴)) _nO)-, wherein, n is 1 to about 6; Or

(b) represent in the following formula two, each is independently selected from :-X ,-R ⁴With-OR ¹

Each R ⁴All be independently selected from H and R ⁵

X represents fluorine, chlorine, bromine or iodine;

Condition is that C15 does not link to each other with Sauerstoffatom.

In preferred compositions: C17 is replaced by alkyl; As C ₁-C ₇Alkyl; Or suc as formula=C (R ⁴) (R ⁴) olefin group, wherein, preferred R ⁴Be hydrogen or C ₁-C ₁₀Alkyl; In preferred embodiments, C17 alkyl eliminating-CH (CH ₃) (CH ₂) ₃CH (CH ₃) ₂In other preferred compositions, C17 is replaced by two atoms that are independently selected from hydrogen and halogen atom; Or C17 is replaced by at least one Sauerstoffatom; Or C17 is replaced by the hydroxyl of hydroxyl or protection; Or C17 is by the carbonyl substituted of carbonyl or protection; Or the C17 alkoxy replaces.In preferred compositions, the substituting group on C17 is got rid of

In other preferred compositions, C15 is replaced by two hydrogen atoms; And/or C4 by hydrogen and-X ,-R ⁴Or-OR ¹One of replace; And/or C5 is replaced by hydrogen; And/or C4 links to each other with at least one hydrogen atom, thereby when C4 linked to each other with two hydrogen atoms, then C3 was not with Sauerstoffatom or do not link to each other with two hydrogen atoms.In another preferred compositions, only to work as C3 and do not link to each other with Sauerstoffatom, when also not linking to each other with two hydrogen atoms, C4 links to each other with two hydrogen atoms.In another preferred compositions, only when C4 did not link to each other with two methyl or formyl radical, C4 linked to each other with methyl.In other preferred compositions, compound has one and be the hydrogen of α configuration on the C5 position.In another preferred compositions, compound have one on the C6 position for the α configuration-OR ¹Group.In another preferred compositions, compound have one on the C7 position for beta comfiguration-OR ¹Group.In another preferred compositions, compound have one on the C6 position for the α configuration-OR ¹Group, have one on the C7 position for beta comfiguration-OR ¹Group.In another preferred compositions, in the compound C3 and C4 at least one link to each other with Sauerstoffatom, in preferred embodiments, C3 links to each other with Sauerstoffatom with C4.In another preferred compositions, the C10 of compound is by methyl substituted; And/or the C13 of compound is by methyl substituted; Or the C10 of compound and C13 are by methyl substituted.In preferred compositions, C6 all links to each other with hydrogen atom with C7.In another preferred compositions, one of C1, C2, C3, C4, C5, C8, C9, C10, C11, C12, C13, C14, C15, C16 and C17 are only replaced by hydrogen atom at least, and more preferably C1 and C2 are only replaced by hydrogen atom; And/or C11 and C12 are only replaced by hydrogen atom; And/or C15 and C16 are only replaced by hydrogen atom.In preferred compositions, compound has saturated A ring; And/or saturated B ring; And/or saturated C ring; And/or saturated D ring.The composition that preferably has saturated A cyclic cpds more preferably has the composition of whole saturated A, B, C and D cyclic cpds.In another preferred compositions, the A of compound ring does not comprise twin nuclei.In another preferred compositions, the C3 of compound and C4 are also only replaced by hydrogen atom together frequently.These compositions can be used for treating asthma, allergy, comprise the inflammation and the thrombosis of sacroiliitis.These compositions also can be mixed with medicine, and the gained medicine for example can be used for treating asthma, allergy, comprises the inflammation and the thrombosis of sacroiliitis.

These compositions for example can be used as analytical standard, a large amount of transportations make things convenient for form or pharmaceutical composition.But the amount of the amount of analysis of The compounds of this invention for being easy to measure according to standard test method well known to those skilled in the art and technology.In the weight of whole composition, but the amount of analysis of The compounds of this invention is generally the about 80wt% of about 0.001-.Inert support comprise any non-degradable or not can with the material of compound generation covalent reaction of the present invention.The suitable example of inert support is a water; Water-containing buffering liquid, as be generally used for that high performance liquid chromatography (HPLC) analyzes those; Organic solvent is as acetonitrile, ethyl acetate, hexane etc.; With pharmaceutically acceptable carrier.

Therefore, the invention provides a kind of medicine or animal medicinal composition (below, abbreviate pharmaceutical composition as), it comprises aforesaid 6,7-titanium dioxide steroide and pharmaceutically acceptable carrier.The present invention also provides a kind of pharmaceutical composition, and it comprises the foregoing 6 of significant quantity, 7-titanium dioxide steroide and pharmaceutically acceptable carrier.

Pharmaceutical composition of the present invention can be any form that composition is taken by the patient.For example, composition can be solid, liquid or gas (aerosol) form.General medication includes but not limited to: oral, topical, parenteral administration, sublingual administration, rectal administration, vagina administration, intranasal administration.Term parenteral administration herein comprises subcutaneous injection, intravenous injection, intramuscular injection, breastbone inner injection or instillation.Pharmaceutical composition of the present invention should be mixed with and make the activeconstituents that is contained in wherein have bioavailability after making composition deliver medicine to the patient.Composition to patient's administration can adopt dose or multidose unit, and for example, tablet can be single dose unit, and can hold multidose unit with the capacity of the steroide of aerosol form.

The material that is used for pharmaceutical compositions should be pharmaceutical purity and nontoxic under the amount that is adopted.The optimal dose that those skilled in the art will appreciate that activeconstituents in the pharmaceutical composition will depend on multiple factor.Correlative factor includes but not limited to: specific form, the administering mode of patient's type (as the mankind), activeconstituents and the composition that is adopted.

Usually, pharmaceutical composition comprises foregoing active 6,7-titanium dioxide steroide and one or more carriers.Described carrier can be particulate state, thereby makes composition can form for example tablet or powder form.If composition is made oral syrup or injectable liquids, then carrier can be liquid.In addition, carrier can be gaseous state, so that provide a kind of aerosol composition to be used for for example inhalation.

When planning oral administration, best solid of composition or liquid form, semisolid, semiliquid, suspension and gel form all are included in the scope that this paper is considered as solid or liquid form.

As the solids composition that is used for oral administration, composition can be mixed with forms such as powder, particle, compressing tablet, pill, capsule, chewing gum, wafer.This solids composition will comprise one or more inert diluents or edible carrier usually.In addition, also can add one or more following assistant agents: tackiness agent such as carboxymethyl cellulose, ethyl cellulose, Microcrystalline Cellulose or gelatin; Vehicle such as starch, lactose or dextrin; Disintegrating agent such as alginic acid, sodiun alginate, Primogel, W-Gum etc.; Lubricant such as Magnesium Stearate or Sterotex; Antiseize paste such as colloidal silica; Sweeting agent such as sucrose or asccharin; Seasonings such as peppermint, wintergreen oil or orange essence, and tinting material.

When composition was capsule such as gelatine capsule, except above-mentioned substance, composition can comprise a kind of liquid vehicle such as polyoxyethylene glycol or lipid acid.

Composition can be liquid form, as elixir, syrup, solution, emulsion or suspension agent.Liquid can pass through oral administration or drug administration by injection.When preparing oral administration, except compound of the present invention, composition preferably comprises one or more sweeting agents, sanitas, dyestuff and/or tinting material and flavour enhancer.Preparing to comprise one or more tensio-active agents, sanitas, wetting agent, dispersion agent, suspension agent, buffer reagent, stablizer and isotonic agent by in the composition of drug administration by injection.

The composition of liquid medicine of the present invention of solution, suspension or other similar type can comprise one or more following assistant agents: sterile diluent such as water for injection, salt brine solution, preferred physiological saline, Ringer's solution, isotonic sodium chloride, and the expressed oil that can be used as solvent or suspension medium is as synthetic single acid glycerol acid or Diglyceride, polyoxyethylene glycol, glycerine, propylene glycol or other solvent; Sterilant such as phenylcarbinol or methyl p-hydroxybenzoate; Antioxidant such as xitix or sodium bisulfite; Sequestrant such as ethylenediamine tetraacetic acid (EDTA); Buffer reagent such as acetate, Citrate trianion or phosphoric acid salt; Regulate reagent such as the sodium-chlor or the glucose of osmotic pressure.The parenteral preparation can be encapsulated in the ampoule.Disposable syringe or multidose bottle can be made by glass or plastics.Physiological saline is preferred assistant agent.It is aseptic that medicinal composition for injections is preferably.

The liquid composition of parenteral administration or oral administration should comprise a certain amount of The compounds of this invention, thereby obtains suitable dosage.Usually, with composition weight meter, described content is at least the The compounds of this invention of 0.01wt%.When oral administration, with composition weight meter, described content is the about 70wt% of 0.01-.The preferred oral composition comprises the active steroide of the about 50wt% of about 4-.Preparing preferred composition of the present invention and preparation makes parenteral dosage unit comprise the active compound of 0.01-1wt%.

But the pharmaceutical composition topical, in this case, carrier can suitably comprise solution, emulsion, ointment or gel matrix.For example, gel matrix can comprise one or more following compositions: Vaseline, lanolin, polyoxyethylene glycol, beeswax, mineral oil, thinner such as water and ethanol, emulsifying agent and stablizer.The pharmaceutical composition that is used for topical also can comprise thickening material.If be intended for use transdermal administration, composition can comprise a kind of percutaneous plaster or ion infiltrates device.During topical, the concentration of The compounds of this invention can be about 0.1-10% (w/v) (weight of per unit volume).

Composition of the present invention can be used for rectal administration, and for example with the suppository form administration, described suppository can discharge medicine in the internal rectum fusing.The composition that is used for rectal administration can comprise a kind of buttery matrix as suitable nonirritant excipient.This matrix includes but not limited to: lanolin, theobroma oil and polyoxyethylene glycol.

Composition of the present invention can comprise the various materials that improve solid or liquid dosage unit physical form.For example, composition can comprise the material that can form coatings around activeconstituents.The material that forms coatings generally is an inert, can be selected from for example sugar, lac and other casing agent.In addition, activeconstituents can be encapsulated in the gelatine capsule.

Thereby the composition of solid or liquid form can comprise and combines the reagent that helps active ingredient to transmit with active steroide component.The suitable reagent that can be used for this purpose comprises mono-clonal or polyclonal antibody, protein or liposome.

Pharmaceutical composition of the present invention can be made up of the gas dose unit, and for example, it can be aerosol form.The term aerosol is used to represent the system of the system of colloidal state to the pressurized package formation.Transmission can distribute the pump system of activeconstituents to finish by liquefied gas or pressurized gas or by a kind of suitable being used to.The aerosol of The compounds of this invention can a phase, two-phase, three-phase system transmit to transmit activeconstituents.Aerocolloidal transmission comprises necessary container, activator, valve and capacitance divider etc., and they can be combined in and form complexes together.Preferred aerosol can be determined by those skilled in the art, need not to carry out any experiment.

No matter be solid, liquid or gas form, pharmaceutical composition of the present invention all can comprise one or more known asthma, allergy, inflammation (comprising sacroiliitis) or thrombotic pharmacologically active reagent of being used for the treatment of.

Pharmaceutical composition can prepare according to the pharmaceutical field known method.Various steroides have been widely used as the activeconstituents in the pharmaceutical composition that is used for the treatment of purposes at present, and according to well known to a person skilled in the art technology, they are also known in the preparation said composition.Steroide of the present invention can be mixed with pharmaceutical composition in a similar fashion.

Plan is to make 6 by the preparation of compositions process of drug administration by injection, 7-titanium dioxide steroide and hydration and and formation solution.Tensio-active agent be can add and uniform solution or suspension are beneficial to form.Tensio-active agent is the compound of covalent effect can not take place with steroide, thereby is convenient to dissolving or the evenly suspension of steroide in moisture transmission system.

Above-claimed cpd and composition have practicality in treatment allergy and asthma, sacroiliitis and/or thrombosis.Aforesaid compound and composition also can be used for treatment and NF _KThe illness that the B value raises and is correlated with wherein, is taken the patient who needs treatment and is reduced NF _KThe compound (or composition of inclusion compound) of the active effective level of B.Herein, " treatment irritated and asthma, sacroiliitis and/or thrombosis " be meant and not only can be used for treatment irritated and asthma, sacroiliitis and/or thrombosis, and can be used for Ammonium Glycyrrhizate reaction, bronchoconstriction, inflammation and the clot formation that causes owing to thrombosis and the development of associated conditions.Herein, NF _KThe B activity is meant directly or indirectly owing to NF _KThe proteinic arbitrary member of B family and all active increase of dna sequence dna bonded genetic transcription or reduction by this family's protein identification.

The compound of significant quantity of the present invention or composition can be used for treating the warm blooded animal such as the mankind's allergy and asthma, sacroiliitis or thrombosis.The method that the anti-allergic agent of significant quantity, anti-asthmatic agent, arthritis agent and antithrombotic agent carry out administration is well known in the art, comprises inhalation, oral administration or parenteral administration.This dosage form includes but not limited to: parenteral solution, tablet, capsule, slowly-releasing implant and transdermal delivery system; Perhaps the inhalation dose system adopts Foradil Aerolizer formoterol fumarate or pressurization multiple doses suction apparatus.Usually, oral or intravenous injection is to be preferred for treatment of arthritis and thrombotic mode.And oral or suction/intranasal administration is for being preferred for treating asthma and mode hypersensitive.Dosage and frequency of utilization all can be selected, and to produce the effective dose of reagent, do not have deleterious effect simultaneously.Usually, when oral administration or intravenous administration were used for antianaphylaxis, anti-asthma, arthritis or antithrombotic treatment, dosage range was about 0.1-100mg/kg/ days, preferably about 0.1-10mg/kg/ days.Equally, when treating to be used for antianaphylaxis and anti-asthma by suction or intranasal administration, dosage range is generally about 0.01-1mg/kg/ days.

When The compounds of this invention or composition are carried out administration, can carry out administration to other medicament simultaneously.For example, wish to give bronchodilator or glucocorticosteroid reagent, give glucocorticosteroid reagent, perhaps give antihistaminic and be used for the treatment of allergy with treatment of arthritis with treatment asthma.Non-steroids can be with steroide co-administered of the present invention, and/or non-steroids can be used for being used in combination so that a kind of one or more methods of treatment of asthma, allergy, sacroiliitis and thrombosis that is used for to be provided with steroide of the present invention.

Following embodiment is used for the present invention is further specified, but they do not limit the present invention.

Except as otherwise noted, flash chromatography method and column chromatography can use Merck silica gel 60 (230-400 order) to finish.Flash chromatography can carry out according to the described process of following document: " purifying of laboratory chemicals ", the 3rd edition, Butterworth-Heinemann Ltd., Oxford (1988) is edited by D.D.Perrin and W.L.F.Armarego, the 23rd page.Column chromatography is meant that eluent flow rate by filler is by the method for gravity decision.Under all scenario, flash chromatography method and radial chromatography can be used alternatingly.Radial chromatography adopts Chromatotron Model#7924T, and (California) silica gel on carries out for Harrison Research, Palo Alto.

The last handling process that routine is used for reaction mixture relates to organic solution (ethyl acetate or ether) diluted reaction mixture, washs organic mixture with saturated sodium bicarbonate, washs with saturated sodium-chloride again.Then, the organic layer dried over mgso is filtered mixture, under the vacuum filtrate is evaporated to driedly, obtains crude product, and crude product can be further purified or not carry out purifying.

The last handling process that routine is used for Wittig reaction at first relates to dropping water so that reaction stops.Then, mixture is diluted with ethyl acetate,, wash with sodium-chlor again with the saturated sodium bicarbonate washing.The organic layer dried over mgso is filtered, and is evaporated to dried.

The last handling process that routine is used for hydroboration relates to pours reaction mixture into saturated nacl aqueous solution (200ml), uses CH ₂Cl ₂Aqueous slurry is extracted, and then, the organic layer after the sodium thiosulfate solution with 25% is combined washs.Use the dried over mgso organic layer again, filter, be evaporated to dried.

Reaction process can adopt thin-layer chromatography (TLC) to monitor usually, adopts silica gel 60 F ₂₅₄Plate (EM Science, Gibbstown, New Jersey) and The suitable solvent system.Tlc can be carried out according to the described process of following document: " purifying of laboratory chemicals ", the 3rd edition, Butterworth-Heinemann Ltd., Oxford (1988) is edited by D.D.Perrin and W.L.F.Armarego, the 30th page.After finishing wash-out, with TLC plate drying, slightly spray 10% vitriolic ethanolic soln, heat then, occur until spot corresponding to compound.Except as otherwise noted, filtration procedure adopts Whatman (I type) filter paper.

Embodiment

Part 1

3,4,6, the polyhydroxylated steroide of 7-synthetic

Having steroide identical or very relevant hydroxylation form with compound 237 (shown in the following structural formula) can begin to synthesize from multiple steroid precursor, and described precursor comprises 4-androstene-3, and 17-diketone (1) and other have C3 oxygen functional group and Δ ⁵The compound of carbon-to-carbon double bond such as dehydroepiandros-sterone (247). 237 (3 β, 4 α, 6 α, 7 β, 17 β) are R wherein ¹=R ³=R ⁴=H, R ²=R ⁵=OH

For example, with androstene-3, the ketone functional group of 17-diketone (1) (embodiment 1, reaction scheme 55) is introduced C7 oxygen with any one protection of many suitable carbonyl-protection bases and simultaneously with after the carbon-to-carbon double bond migration by the allylic oxidation.This oxidation step can use multiple oxygenant and experiment condition, includes but not limited to that chromium trioxide/3 title complex, chloro chromic acid pyridine (PCC), dichromic acid pyridine (PDC) and tertbutyl peroxide add RuCl ₃The ketone that generates is generated the hydroxyl functional base with suitable reductive agent reduction in the C7 position.This process can be used multiple metal hydride, comprises sodium borohydride and lithium aluminium hydride.Usually, the reduction of C7 carbonyl generates β OH configuration by hydride from a side attack of steroidal steric hindrance minimum.

With the C7 hydroxyl with suitable protecting group protection after, can introduce C6 oxygen by for example hydroboration/oxidizing reaction or epoxidation and open loop subsequently.Can be with Δ ⁵Carbon-to-carbon double bond comprises metachloroperbenzoic acid, trifluoroperacetic acid or 3 with any one peracid epoxidation, described peracid, 5-dinitrobenzene benzoyl hydroperoxide.Usually, the epoxide of introducing has the α configuration, and this is owing to the side attack from steroidal ring structure steric hindrance minimum forms.Can for example in 80% aqueous acetic acid, under 60 ℃, make the epoxide open loop at acidic conditions subsequently.Generate vinyl carbinol thus with α configuration in the C6 position.Perhaps, with Δ ⁵Two keys also also can be introduced the hydroxyl of α configuration subsequently in the C6 position with reagent oxidation such as alkaline hydrogen peroxides with suitable borane complexes hydroboration.

Can encircle 4-alkene-3-ketone functional group form from A-begins to introduce hydroxyl at C3 and C4 position.The available lithium that is dissolved in liquefied ammonia of the reduction of alpha, beta-unsaturated ketone is finished.The enolate that generates can be caught with electrophilic reagent such as trimethylsilyl chloride or chloro diethyl phosphoric acid.The silylation enol ether is carried out hydroboration-oxidizing reaction and introduce oxygen at C4.This method generates 3 β, 4 α hydroxyl configurations.Perhaps, enol phosphate is carried out the reduction second time with the lithium in the liquefied ammonia and produce Δ ³Carbon-to-carbon double bond.With Δ ³Two keys generate α-epoxide with peracid such as metachloroperbenzoic acid epoxidation.Can adopt multiple acidity or alkaline condition to make this epoxide open loop.For example, with 3 α in the compound 238 (embodiment 3, reaction scheme 61), 4 α-epoxide function base is handled with glacial acetic acid can generate 3 Alpha-hydroxies, 4 β-acetoxyl group form.In methyl alcohol, remove ethanoyl with any one reagent such as salt of wormwood (or sodium methylate) and generate 3 α, 4 beta-hydroxy forms.

Embodiment 1

Steroidal 3 β, 4 α, 6 α, 7 β, 17 β-penta hydroxy group-5 α-etioallocholane (237)

Can be synthetic according to the reaction sequence shown in the reaction scheme 55

Reaction scheme 55

Annotate: (ⅰ) p-TsOH, (CH ₂OH) ₂, benzene (ⅱ) CrO ₃, 3, methylene dichloride (ⅲ) NaBH ₄, CeCl ₃, THF-MeOH (ⅳ) TBDMSCl, imidazoles, DMF (ⅴ) m-CPBA, methylene dichloride (ⅶ) 80%AcOH (ⅶ) TBAF, THF (ⅷ) 2,2-Propanal dimethyl acetal, CSA, DMF (ⅸ) NaBH ₄, MeOH (ⅹ) TBDMSCl, imidazoles, DMF (ⅹ ⅰ) 1.Li/NH ₃-THF, 2.TMSCl, Et ₃N (ⅹ ⅱ) 1.BH ₃-THF title complex, 2.30%NaOH, 30%H ₂O ₂, THF (ⅹ ⅲ) TBAF, THF (ⅹ ⅳ) p-TsOH, H ₂O, THF.

Under the condition of refluxed under nitrogen, the 4-androstene-3 that can buy, 17-diketone (1) (20.0g, 62.8mmol) and the tosic acid of ethylene glycol (10ml) and catalytic amount (1.0g 5.2mmol) stirs 26 hours (reaction scheme 56) together in benzene (500ml).During this period, remove the water that dereaction generated with Dean Stark equipment.Then mixture is cooled to room temperature and adds ether.Mixture washs with sodium bicarbonate, washes and use dried over mgso then with water.Filter and concentrate and obtain light yellow solid, it is obtained two ketals 2 (14.6g, 39.0mmol, 62%) of white powder with methanol wash.From filtrate, reclaim the single ketal by product (5.13g, 15.0mmol) and recycle.Consider by product, the total recovery of two ketals is 86%.

In methylene dichloride, the allylic oxidation is carried out in the C7 position of two ketals 2 with chromium trioxide-3 title complex and generate compound 3 (reaction scheme 56).Under the nitrogen, (46.7g 467mmol) and anhydrous methylene chloride (450ml), is cooled to-20 ℃ with dry ice-calcium chloride solution then to add chromium trioxide in flask.(44.9g 467mmol) and with this mixture stirred 1.5 hours in-20 to-30 ℃ to add 3.(7.00g 18.7mmol) and at-20 ℃ continues down to stir 7 hours to add two ketals 2 subsequently.The water termination reaction is also filtered.With the filtrate water washing, volume is concentrated into 200ml uses dried over mgso then.Filter and concentrate and obtain dark-brown oil, it is carried out purifying with silica gel column chromatography (dichloromethane/ethyl acetate) obtain ketenes 3, and yield 68% (4.95g, 12.8mmol).

In THF and methyl alcohol, ketenes 3 is reduced into vinyl carbinol 4 (reaction scheme 56) with sodium borohydride and seven hydration Cerium II Chlorides (III).Under the nitrogen, to contain ketenes 3 (15.3g, 39.4mmol) and CeCl ₃7H ₂(16.0g adds new THF (200ml) and the methyl alcohol (50ml) that steams to O in flask 42.9mmol).Add sodium borohydride (3.20g 84.6mmol), continues to stir 1 hour under room temperature then in batches.Add methylene dichloride, mixture is used sodium hydroxide solution (0.6N) and water washing successively, then with the organic layer dried over mgso.Filter and concentrate and obtain white solid compound 4 (14.1g, 36.1mmol, 92%).

Reaction scheme 56

With the form of silyl ether the C-7 hydroxylic moiety of alcohol 4 is protected (reaction scheme 57) then.With compound 4 (14.1g 36.1mmol) is dissolved in dry DMF (50ml), add then imidazoles (5.9g, 86.7mmol) and TBDMSCl (6.7g 44.5mmol) and with this mixture stirred 5 hours down in nitrogen.Add methylene dichloride, mixture is washed with water the organic phase dried over mgso.Filter and concentrate and obtain faint yellow solid, it is obtained white solid compound 5 with recrystallizing methanol, and yield 61% (11.2g, 22.2mmol).

Use metachloroperbenzoic acid (m-CPBA) with compound 5 epoxidations (reaction scheme 57) subsequently.(0.72g 1.4mmol) is dissolved in anhydrous methanol (5ml), and (0.50g is 2.9mmol) and with mixture vigorous stirring 20 minutes to add m-CPBA with compound 5.Add saturated sodium bicarbonate solution also with this aqueous slurry dichloromethane extraction.The organic extract liquid that merges is used sodium carbonate solution, water, 10% Sulfothiorine and water washing successively.Dry (sal epsom), filter and concentrate and obtain white solid, it is carried out purifying with flash chromatography obtains compound 6, yield 77% (0.57g, 1.1mmol).

With compound 6 usefulness acid treatment with C3 and C17 ketone deprotection and make the epoxide moiety open loop generate 6-hydroxyl-7-siloxy-compound 310 (reaction scheme 57).In the flask that contains compound 6, add acetic acid aqueous solution (80%, 1ml).Compound in 65 ℃ of heating 5 hours, is cooled off and pours in the methylene dichloride.Compound is washed and uses dried over mgso with sodium hydrogen carbonate solution.After filtering and concentrating, be not directly used in next step reaction with the crude product 310 that obtains is purified.

Under room temperature and the nitrogen, (2.30mg, THF 5.32mmol) (10ml) solution is handled 10 minutes to remove silyl protecting group (reaction scheme 57) with tetrabutyl ammonium fluoride (TBAF) (8ml, the THF solution of 1M) with compound 310.Reaction mixture is concentrated, then by flash chromatography (3: 1 dichloromethane/ethyl acetate) carry out purifying obtain compound 7 (1.37mg, 4.31mmol), yield 81%.

Reaction scheme 57

By with 2, (1S)-(+) of 2-Propanal dimethyl acetal and catalytic amount-10-camphorsulfonic acid (CSA) is handled 6 of compound 7, and the 7-glycol is protected, and generates acetone solvate 8 (reaction scheme 58).(1g, 3.14mmol) CSA with catalytic amount is dissolved in dry DMF (2ml) and 2,2-Propanal dimethyl acetal (10ml) with compound 7.Mixture was heated 0.5 hour down in 100 ℃.Adding methylene dichloride also washes this mixture with water.With the organic layer dried over mgso, filter and concentrate and obtain compound 8 (1.10g, 3.07mmol, 98%), this product is not purified to be directly used in next step reaction.

The C-17 carbonyl moiety is carried out chemical selective reduction (reaction scheme 58), and the alcohol that will generate is as required protected with the form of silyl ether subsequently.(83mg 0.23mmol) is dissolved in methyl alcohol (250ml) and in batches added sodium borohydride (15mg) in 1.5 hours under nitrogen with compound 8.After 30 minutes, use the acetate termination reaction, neutralize with sodium bicarbonate then.Steam and remove methyl alcohol, resistates is recovered in the methylene dichloride.Mixture is washed and uses with water dried over mgso.Obtain primary product compound 9 (72mg, 0.20mmol, 87%) through flash chromatography (2: 1 dichloromethane/ethyl acetate).

The C17 hydroxyl is protected generation compound 10 with the form of silyl ether, with liquefied ammonia lithium-THF the alpha, beta-unsaturated ketone in the A-ring is reduced subsequently, catch enolate (reaction scheme 58) with trimethylsilyl chloride.Under-78 ℃, (75.2mg, 0158mmol) solution in the trimethyl carbinol (0.020ml) and THF (1.5ml) is transferred to the anhydrous liquid ammonia solution that contains metallic lithium (11.4mg is in flask 10ml) with compound 10.-78 ℃ after following 20 minutes, add isoprene (0.5ml) to destroy excessive lithium.Then this mixture being warming up to the steaming of room temperature and vacuum desolventizes.Resistates is dissolved in THF (5ml), be cooled to-78 ℃ and add triethylamine (1.1ml, 0.30mmol) and TMSCl (0.80ml, 0.30mmol).Remove cooling bath and mixture was stirred 15 minutes.Add saturated sodium bicarbonate and with aqueous layer with diethyl ether and dichloromethane extraction.With the organic layer salt solution washed twice that merges, with dried over mgso and concentrated.Crude product is obtained compound 235 by the radial chromatography purifying, and yield 67% (58.5mg, 0.10mmol).

By C4 hydroxyl (reaction scheme 58) is introduced in silyl enol ether 235 hydroborations-oxidation.(58.5mg 0.106mmol) is dissolved in anhydrous THF (15ml) and cooling off in ice bath with compound 235.(1.0M THF title complex: 0.32ml 0.32mmol), is warming up to room temperature with this mixture and stirred 45 minutes to add borine.Add BH once more ₃-THF title complex (0.16ml) also continues to stir 2 hours.Then mixture is cooled off in ice bath and add 15%NaOH (0.5ml) and 30%H ₂O ₂(0.5ml).Continued vigorous stirring 2 hours.Then water layer is used methylene dichloride and extracted with diethyl ether successively, the organic extract liquid that merges is used 1O%Na successively ₂S ₂O ₃The aqueous solution and salt water washing are also used dried over mgso.Crude product is carried out purifying by radial chromatography obtain compound 236 (34.0mg, 0.0688mmol, 65%) and corresponding 3 β-silyl ether (11.8mg, 0.0208mmol, 20%).

Compound 236 earlier with TBAF deprotection in THF, is used aqueous acid-THF deprotection then, generate 3 β, 4 α, 6 α, 7 β, 17 β-penta hydroxy group-5 α-etioallocholane (237).

Reaction scheme 58

Embodiment 2 steroidals 3 α, 4 α-epoxy-6 α, 7 β, 17 β-trihydroxy--5 α-etioallocholane (239)

Can be synthetic according to the reaction sequence shown in the reaction scheme 59

Reaction scheme 59

Annotate: (ⅰ) p-TsOH, (CH ₂OH) ₂, benzene (ⅱ) CrO ₃, 3, methylene dichloride (ⅲ) NaBH ₄, CeCl ₃, THF-MeOH (ⅳ) TBDMSCl, imidazoles, DMF (ⅴ) m-CPBA, methylene dichloride (ⅵ) 80%AcOH (ⅶ) TBAF, THF (ⅷ) 2,2-Propanal dimethyl acetal, CSA, DMF (ⅸ) NaBH ₄, MeOH (ⅹ) TBDMSCl, imidazoles, DMF (ⅹ ⅰ) 1.Li/NH ₃-THF, 2.Cl (O) P (OEt) ₂(ⅹ ⅱ) Li/NH ₃, the trimethyl carbinol (ⅹ ⅲ) m-CPBA, methylene dichloride (ⅹ ⅳ) TBAF, THF (ⅹ ⅴ) p-TsOH, THF, H ₂O.

3 α, 4 α-epoxy-6 α, 7 β, 17 β-trihydroxy--5 α-etioallocholane (239) can be from synthetic 3 β, 4 α, 6 α, 7 β, the intermediate 10 of 17 β-penta hydroxy group-5 α-etioallocholane (258) (reaction scheme 60) makes.Under-78 ℃ and the argon gas, (111mg, THF 0.234mmol) (4ml) solution transfer to and contain the metallic lithium liquid ammonia solution (6.4mg is in flask 10ml) with compound 10.-78 ℃ after following 30 minutes, add isoprene (0.5ml) to destroy excessive lithium.Mixture is warming up to room temperature and vacuum to be steamed and desolventizes.Resistates is dissolved in THF (5ml), is cooled to-78 ℃, add Cl (O) P (OEt) ₂(0.044ml 0.30mmol) and with mixture stirred 1 hour.Add entry and methylene dichloride, with the mixture acidifying, with water layer methylene dichloride and extracted with diethyl ether.Wash and use the organic layer that merges with water dried over mgso.Crude product is obtained compound 11 by the radial chromatography purifying, and yield 59% (85.1mg, 0.139mmol).

The liquid ammonia solution of compound 11 usefulness lithiums reduced in the presence of the trimethyl carbinol generate alkene 128.Under-30 ℃ and the argon gas, with compound 11 (85.1mg, 0.139mmol) and THF (6ml) solution of the trimethyl carbinol (0.05ml) transfer in the flask of the liquid ammonia solution that contains metallic lithium (16mg).After 30 minutes, make the ammonia evaporation, add entry then.Behind hcl acidifying, mixture is used ether and ethyl acetate extraction successively, wash and use the organic layer that merges with water dried over mgso then.Crude product is obtained compound 128 by the radial chromatography purifying, and yield 78% (50.3mg, 0.109mmol).

128 usefulness metachloroperbenzoic acids (m-CPBA) epoxidation in methylene dichloride is generated epoxide 238.(50.3mg 0.109mmol) is dissolved in anhydrous methylene chloride (1.5ml) and add m-CPBA (43.0mg) with compound 128.With mixture stirring at room 1.5 hours, transfer in the separating funnel then and use 10%Na ₂S ₂O ₃, saturated sodium bicarbonate solution and water washing, use dried over mgso.Filter and evaporated filtrate (52.0mg 0.109mmol) obtains compound 228, and this product is not purified to be directly used in next step reaction with 78% yield.

Compound 238 earlier with TBAF deprotection in the THF that refluxes, is generated compound 239 with acid THF aqueous solution deprotection then.

Reaction scheme 60

Embodiment 3 steroidals 3 α, 4 β, 6 α, 7 β, 17 β-penta hydroxy group-5 α-etioallocholane (241) can be synthetic according to following reaction sequence (reaction scheme 61)

Reaction scheme 61 Annotate: (ⅰ) p-TsOH, (CH ₂OH) ₂, benzene (ⅱ) CrO ₃, 3, methylene dichloride (ⅲ) NaBH ₄, CeCl ₃, THF-MeOH (ⅳ) TBDMSCl, imidazoles, DMF (ⅴ) m-CPBA, methylene dichloride (ⅵ) 80%AcOH (ⅶ) TBAF, THF (ⅷ) 2,2-Propanal dimethyl acetal, CSA, DMF (ⅸ) NaBH ₄, MeOH (ⅹ) TBDMSCl, imidazoles, DMF (ⅹ ⅰ) 1.Li/NH ₃-THF, 2.Cl (O) P (OEt) ₂(ⅹ ⅱ) Li/NH ₃-THF, the trimethyl carbinol (ⅹ ⅲ) m-CPBA, methylene dichloride (ⅹ ⅳ) acetate, 60 ℃ of (ⅹ ⅴ) salt of wormwood, MeOH, (ⅹ ⅵ) TBAF that refluxes, THF refluxes.

Adopt and synthetic 3 α, 4 α-epoxy-6 α, 7 β, the identical condition of 17 β-trihydroxy--5 α-etioallocholane (239) is from intermediate 238 synthetic 3 α, 4 β, 6 α, 7 β, 17 β-penta hydroxy group-5 α-etioallocholane (241).Subsequently by heating the epoxide open loop with glacial acetic acid and removing the compound 146 that acetone solvate functional group (reaction scheme 62) obtains containing at C4 the acetoxyl group functional group simultaneously.(18.5mg adds acetate (0.3ml) in flask 0.038mmol) to containing compound 238.With this mixture heating up to 60 ℃ stirring 24 hours, stirring at room was 2 days then.The vacuum steaming removes acetate and obtains compound 146, and yield 93% (18mg, 0.036mmol).

With salt of wormwood in the methyl alcohol that refluxes with C4 hydroxyl deprotection, with TBAF C17 hydroxyl deprotection is obtained pentahydroxyl compound 241 then.

Reaction scheme 62

Embodiment 4

3 β, 4 α, 6 α, 7 β, 17 β-penta hydroxy group-5 α-etioallocholane 6,7-acetone solvate (246)

Steroidal 3 β, 4 α, 6 α, 7 β, 17 β-penta hydroxy group-5 α-etioallocholane 6,7-acetone solvate (246) can be synthetic according to reaction scheme 63 described reaction sequence.

Reaction scheme 63

Annotate: (ⅰ) 80%AcOH (ⅱ) TESCl, imidazoles, DMF (ⅲ) 1.Li/NH ₃-THF, 2.TMESCl, triethylamine (ⅳ) 1.BH ₃-THF, 2.30%NaOH, 30%H ₂O ₂(ⅴ) 2,2-Propanal dimethyl acetal, CSA, DMF (ⅵ) TBAF, THF.

Compound 10 prepares according to the description among part 1, the embodiment 1.,, after appropriate postprocessing, these groups are protected again with the form of silyl ether derivant obtain 242 to 6,7 and 17 hydroxylic moiety deprotections with 80% acetic acid solution.Compound 10 is dissolved in 80% acetate, and stirring at room is this mixture vacuum concentration after 8 hours.Add resistates in the dry DMF contain imidazoles and TBDMSCl and stirring at room 20 hours under nitrogen atmosphere.Adding ether can be with this mixture successively with 5% hydrochloric acid, saturated sodium bicarbonate solution and saturated nacl aqueous solution washing.With the organic mixture dried over mgso, filter and evaporation.Resistates is obtained compound 242 by the silica gel chromatography purifying.

With the alpha, beta-unsaturated ketone reduction in the A-ring, catch enolate with liquefied ammonia lithium-THF with trimethylsilyl chloride.Under-78 ℃, compound 242 is transferred in the flask of the anhydrous liquid ammonia solution that contains metallic lithium at the solution in 1: 4 trimethyl carbinol one THF mixture.-78 ℃ after following 20 minutes, add isoprene (0.5ml) to destroy excessive lithium.Then this mixture being warming up to the steaming of room temperature and vacuum desolventizes.Resistates is dissolved in THF, is cooled to-78 ℃, add triethylamine and TMSCl.Remove cooling bath and mixture was stirred 15 minutes.Add saturated sodium bicarbonate and with water layer with ether and dichloromethane extraction.With the organic layer salt solution washed twice that merges, with dried over mgso and concentrated.Crude product is obtained compound 243 by the radial chromatography purifying.

By the C4 hydroxyl is introduced in silyl enol ether 243 hydroborations-oxidation.Compound 243 is dissolved in anhydrous THF and in ice bath, cools off.Add borine (1.0M THF title complex), this mixture is warming up to room temperature and stirred 45 minutes.Add BH once more ₃-THF title complex also continues to stir 2 hours.Then mixture is cooled off in ice bath and add 30%NaOH and 30%H ₂O ₂Continued vigorous stirring 12 hours.Then water layer is used methylene dichloride and extracted with diethyl ether successively, the organic extract liquid that merges is used 10%Na successively ₂S ₂O ₃The aqueous solution and salt water washing are also used dried over mgso.Crude product is carried out purifying by radial chromatography obtain compound 244.

By with 2, (1S)-(+) of 2-Propanal dimethyl acetal and catalytic amount-10-camphorsulfonic acid (CSA) is handled 2 of compound 244, and the 3-glycol is protected, and generates acetone solvate 245.The CSA of compound 244 and catalytic amount is dissolved in dry DMF and 2, the 2-Propanal dimethyl acetal.Mixture was heated 0.5 hour down in 100 ℃.Adding methylene dichloride also washs this mixture with saturated sodium bicarbonate solution.With the organic layer dried over mgso, filter and concentrate and obtain compound 245, this product is not purified to be directly used in next step reaction.

With TBAF compound 245 is transformed into triol 246 then.Three silyl ethers 245 are dissolved in THF and use tetrabutyl ammonium fluoride (TBFA) (the THF solution of 1M) to handle 5 hours down in room temperature and nitrogen.Reaction mixture is poured in the methylene dichloride and use into the salt water washing, drying (sal epsom) and vacuum concentration.Resistates is obtained compound 246 by flash chromatography (dichloromethane/ethyl acetate) purifying.

Part 2

22,29-epoxy-15-ketone steroid synthetic

With 22,29-epoxy-3,4,6,7,29-penta hydroxy group-14 β-stigmastane-15-ketone (165) steroide relevant, that have the C15 ketone group and have a hemiacetal functional group (being that C22 hydroxyl functional base and the condensation of C29 aldehyde functional group form an amylene oxide ring and C29 hydroxyl) on the steroidal side chain can synthesize by several different methods.Committed step comprises synthetic (in the 3rd part description being arranged) and the coupling of side chain of the introducing of C15 oxygen, suitable side chain.Can use and multiplely commercially available contain the steroide (for example Vitarrine (C17 ethanoyl), 4-androstene-3,17-diketone (C17 ketone), dehydroepiandros-sterone (C17 ketone)) of ketone group or ethanoyl as raw material at C17.

In a kind of method therein (embodiment 5, reaction scheme 64), C17 ketone is handled introducing (Z)-17 (20) ethylidene parts with Witting reagent, described Witting reagent makes by ethyl triphenyl phosphonium bromide and potassium tert.-butoxide are reacted in THF.With Wittig product and compound such as 5-acetoxy-3-(1 '-methylethyl) valeral (156 of containing aldehyde; the 3rd part; embodiment 8; reaction scheme 71) the reaction generation contains the required C22 oxygen product of (form with the hydroxyl functional base exists) in the presence of Lewis acid, and C29 oxygen is with the form protection of acetic ester.This olefination is created on two kinds of isomer of C22, and the ratio of isomer depends on the selection of reaction conditions and raw material aldehyde.Therefore, 3R by adopting compound 156 or 3S isomer can synthesize at C22 and C24 and have not four kinds of possible diastereomers of isomorphism type.

Above-mentioned olefination generates Δ ¹⁶Carbon-to-carbon double bond, this pair key can be used to introduce C15 oxygen by the allylic oxidizing reaction.For example, after the protecting group such as t-butyldimethylsilyl protection that the C22 hydroxyl usefulness of alkylene product is suitable, carry out the allylic oxidizing reaction, introduce the ketone functional group in the C15 position with chromium trioxide/reagent such as 3 title complex.With Δ ¹⁶The carbon-to-carbon double bond reduction generates and 22,29-epoxy-3,4,6,7, and 29-penta hydroxy group-14 β-stigmastane-15-ketone (165) has the steroide of identical D-ring function base.

Remove C29 protecting group (being generally acetate groups), subsequently the primary alconol oxidation that generates is formed required aldehyde at C29.Remove C22 protecting group (being generally t-butyldimethylsilyl) form with compound 165 in identical side chain hemiacetal part.

The second kind of scheme that generates required hemiacetal side chain and C15 ketone functional group relates to introduces C15 oxygen (embodiment 6, reaction scheme 67) before side chain coupling.In the method, the first step comprises, changes the acetate enol ester into by the C17 ketone functional group of handling the steroide intermediate with methylvinyl acetate and tosic acid.Handle with acid chloride and tributyl tin methylate then and change the acetate enol ester into alpha, beta-unsaturated ketone.

Use then from p-methoxybenzyl alcohol deutero-alkoxide and in the presence of potassium hydroxide, ketenes is carried out Michael addition introducing C15 oxygen.Can carry out deprotection optionally and the alcohol that will generate is oxidized to C15 ketone to this functional group then.

The process that C17 ketone functional group is transformed into ethanoyl at first is with the attack of acetylene negatively charged ion.For this reason, can use commercially available acetylene lithium-ethylene diamine complex.Product dehydration generation is had the conjugated Δ ¹⁶The compound of carbon-to-carbon double bond.Adopt the reagent such as Dowex resin of for example mercury infiltration in methyl alcohol, THF and water, the acetylene moiety hydration to be generated ethanoyl (Δ at C17 ¹⁶, C20 ketone).With Δ ¹⁶Carbon-to-carbon double bond reduces under condition of phase transition with V-Brite B and sodium bicarbonate.

By using the ylide that among THF, makes from trimethyl sulfonium iodide and n-Butyl Lithium to handle methyl ketone is transformed into epoxide then.Then this epoxide is carried out the Stereoselective open loop with Lewis acid as magnesium bromide ether compound and generate C22 aldehyde.Make the alkyl negatively charged ion by lithium-halogen exchange reaction from suitable halogenide such as iodide 284 (embodiment 9, reaction scheme 72), then with the side chain of protection form identical in this alkyl negatively charged ion attack steroide aldehyde generation and the compound 165.C29 aldehyde deprotection is obtained required side chain.

Provided the detailed content of above two kinds of schemes among the embodiment 5 and 6.

Embodiment 5

22,29-epoxy-3,29-dihydroxyl-14 β-stigmastane-15-ketone (260)

The example of the first method of introducing as part 2, steroidal 22,29-epoxy-3,29-dihydroxyl-14 β-stigmastane-15-ketone (260) can be synthetic according to the reaction sequence of describing in the reaction scheme 64.

Reaction scheme 64

Annotate: (ⅰ) H ₂, Pd/C, ethyl acetate (ⅱ) EtPh ₃PBr, potassium tert.-butoxide, THF (ⅲ) TBDMSCl, imidazoles, DMF (ⅳ) 156, Me ₂AlCl, methylene dichloride (ⅴ) TBDMSCl, imidazoles, DMF (ⅵ) CrO ₃, 3, methylene dichloride (ⅶ) H ₂, Pd/C, ethyl acetate (ⅷ) salt of wormwood, methyl alcohol (ⅸ) PCC, sodium acetate, methylene dichloride (ⅹ) TBAF, THF (ⅹ ⅰ) KOH, methyl alcohol.

22,29-epoxy-3, the synthetic of 29-dihydroxyl-14 β-stigmastane-15-ketone (260) can begin to finish by 10 steps from dehydroepiandros-sterone (247).With the Δ in the compound 247 ⁵The carbon-to-carbon double bond catalytic hydrogenation generates the compound 250 that contains the trans A/B of condensing ring system.Compound 247 is dissolved in ethyl acetate and adds 10%Pd/C.With this mixture stirred overnight at room temperature under hydrogen.Obtain compound 250 with diatomite filtration is also concentrated, this product can be directly used in next step reaction.

With the phosphorus ylide that among THF, makes from ethyl triphenyl phosphonium bromide and potassium tert.-butoxide compound 250 is carried out Wittig reaction and obtain compound 251.The form of ethyl triphenyl phosphonium bromide with suspension stirred in THF.Under nitrogen gas stream, add potassium tert.-butoxide and with mixture stirring at room 1 hour.The form of compound 250 with THF solution joined in the formed negatively charged ion.Mixture refluxed under nitrogen 2 hours, is cooled to room temperature then and drips the water termination reaction.Add saturated ammonium chloride solution and with the water layer ethyl acetate extraction, with the organic layer water and the salt water washing of merging and use dried over mgso.Filter and concentrate and obtain crude product 251, it is carried out purifying by flash chromatography on silica gel, carry out the stage gradient wash-out with hexane and ethyl acetate.

In DMF, 3 beta-hydroxy protections are obtained 252 with tert-butyldimethylsilyl chloride and imidazoles.Compound 251 is dissolved in DMF and adds imidazoles.After adding TBDMSCl, with the mixture stirred overnight at room temperature.Add methylene dichloride then and mixture is washed with water, then with the organic phase dried over mgso.Filter and concentrate and obtain crude product 252, this product can not purified direct use.

Then compound 252 and aldehyde 156 coupling in the presence of suitable Lewis acid are obtained on the compound 253.Aldehyde 156 is dissolved in methylene dichloride and adds Me down in-78 ℃ ₂AlCl (hexane solution of 1.0M).After 5 minutes, add the dichloromethane solution of compound 252.Then this mixture was warming up to room temperature in 16 hours.Then it is cooled to-78 ℃ and also uses the methanol/water mixture termination reaction.Layering and with the water extracted with diethyl ether.The organic phase that merges is used 1N hydrochloric acid, saturated sodium bicarbonate aqueous solution and salt water washing successively, use dried over mgso then.Filter and be evaporated to do after, the C22 isomer separated obtaining compound 253a and 253b with flash chromatography on silica gel.

Compound 253a (or 253b) is dissolved in dry DMF and adds imidazoles.Add TBDMSCl and, stirred 3 hours down in 60 ℃ then this mixture stirring at room 14 hours.Mixture is diluted with ether, use dried over mgso after washing with water.Filter and evaporation after, with crude product flash chromatography on silica gel purifying, obtain compound 254 with the mixture wash-out of ethyl acetate and hexane.

In methylene dichloride, carry out the allylic oxidizing reaction at the C15 of compound 254 with chromium trioxide and 3 and obtain compound 255.Join chromium trioxide and methylene dichloride in the flask and be cooled to-20 ℃, under this temperature, stirred 15 minutes then.Add 3, then reaction solution was stirred 1.5 hours.The dichloromethane solution that adds compound 254 is also placed this mixture 5 days down in-20 ℃.Then this mixture is warming up to room temperature and uses filtered through silica gel, wash with ethyl acetate.Steaming desolventizes and obtains crude product, and it is obtained purified compound 255 with flash chromatography (ethyl acetate/hexane) purifying.

Can with hydrogen and palladium charcoal in ethyl acetate with the Δ in the compound 255 ¹⁶The carbon-to-carbon double bond reduction.Compound 255 is dissolved in ethyl acetate and adds the 10%Pd/C of catalytic amount.Mixture stirred under nitrogen atmosphere spend the night,, obtain compound 256 behind the purifying then with diatomite filtration and concentrate.

Available salt of wormwood removes in (or with sodium methylate at methyl alcohol) in methyl alcohol that the acetic ester protecting group on the side chain obtains compound 257 in the compound 256.Compound 256 is dissolved in methyl alcohol and adds salt of wormwood.Mixture was refluxed 3 hours, be cooled to room temperature and pour in the methylene dichloride.Add sodium bicarbonate aqueous solution (10%) and separatory.With the water layer dichloromethane extraction, wash and use the organic extract liquid that merges with water dried over mgso.Filter, evaporation and purifying obtain compound 257.

Available PCC is oxidized to aldehyde 258 with the primary alconol 257 that generates.Compound 257 and sodium acetate stirred in methylene dichloride and add PCC.With mixture stirring at room 3 hours, use diatomite filtration then.Filtrate is concentrated and resistates is obtained compound 258 by the flash chromatography purifying.

Available tetrabutyl ammonium fluoride removes the protecting group of two hydroxylic moieties in the compound 258 in a step simultaneously.Compound 258 is dissolved in THF and adds the THF solution of tetrabutyl ammonium fluoride.With the mixture stirred overnight at room temperature, vacuum concentration and purifying obtain 22 then, 29-epoxy-3,29-dihydroxyl-14 α-stigmastane-15-ketone (248).

The methanol solution of compound 248 usefulness potassium hydroxide is carried out epimerization in the C14 position obtain 22,29-epoxy-3,29-dihydroxyl-14 β-stigmastane-15-ketone (260).Compound 248 is dissolved in methyl alcohol and adds the methanol solution (25mg/ml) of potassium hydroxide.Mixture was refluxed 15 minutes, be cooled to room temperature then.Add entry, with this aqueous slurry with chloroform extraction and use dried over mgso.Filter and concentrate the crude product of the epimer mixture that obtains containing compound 248 and 260.Available column chromatography is separated with 260 248.

Another route of synthetic compound 260 (reaction scheme 65) relates to preparation side chain delta-lactone and carries out the compound that the Dibal-H reduction obtains containing C29 hemiacetal functional group subsequently.For example, 3 beta-hydroxies in the compound 251 are protected with the form of benzyloxy, carried out the olefination of embodiment 5 described routines subsequently.In methyl alcohol, the C29 acetoxyl group is carried out deprotection with sodium methylate then.The silver carbonate oxidation in the benzene that refluxes that then glycol 263 usefulness that generate is adsorbed on the diatomite obtains delta-lactone.Compound 263 is dissolved in benzo adds the silver carbonate that is adsorbed on the diatomite, then mixture was refluxed 12 hours.Reaction mixture is filtered, and evaporation also obtains lactone 264 with resistates by the flash chromatography purifying.Compound 264 usefulness chromium trioxides and 3 are carried out the allylic oxidation in C15 carbonylate part (compound 265) in methylene dichloride.With hydrogen and palladium charcoal in ethyl acetate with the conjugated Δ ¹⁶The carbon-to-carbon double bond reduction is also used sodium methylate to remove the benzoic ether group in the chloroform/methanol at 1: 1 subsequently and is obtained product 266.At last, with of the form protection of C15 ketone, under-78 ℃, delta-lactone optionally is reduced into lactol with DIBAL then and also obtains compound 260 with 80% acetic acid aqueous solution deprotection with ethylene ketal.

Reaction scheme 65 Annotate: (ⅰ) H ₂, Pd/C, 1: 1 ethyl acetate/ethanol (ⅱ) EtPh ₃PBr, potassium tert.-butoxide, toluene (ⅲ) BnBr, NaH, DMF (ⅳ) 156, (CH ₃) ₂AlCl, methylene dichloride (ⅴ) sodium methylate, methyl alcohol (ⅵ) Ag ₂CO ₃/ diatomite, benzene (ⅶ) CrO ₃, 3, methylene dichloride (ⅷ) 1.H ₂, Pd/C, ethyl acetate 2. sodium methylates, methyl alcohol (ⅸ) 1.HOCH ₂CH ₂OH, pTsOH, benzene 2.DIBAL, methylene dichloride ,-78 ℃, 3.80% acetate.

Embodiment 6

As described in the beginning part of this section, the second method of introducing C15 oxygen relates to uses Cantral etc., organic chemistry magazine, 29:64, the Michael reaction described in 1963.Need the selectively removing protecting group and subsequently the secondary alcohol that forms is oxidized to ketone on the C15.Horita etc., tetrahedron (Tetrahedron), 42 (11): 3021-3028, confirm in 1986, right-methoxy-benzyl protecting group can multiple other protecting group comprise benzyl in the presence of with 2,3-two chloro-5,6-dicyano benzoquinone (DDQ) removes.Therefore, among KOH, make alkoxide also with used similar benzyloxy alkoxide such as this alkoxide replaced C antral from p-methoxybenzyl alcohol.Following reaction scheme 66 has provided the example that generates this chemical reaction of ketene compound 270, described compound 270 usefulness Takahashi etc., and tetrahedron, 41 (24): 5747-5754, the method described in 1985 makes from transdehydroandrosterone (247).

Reaction scheme 66

The reaction process of the Michael addition described in the reaction scheme 67 is as follows.Compound 270 is dissolved in p-methoxybenzyl alcohol and adds the potassium hydroxide powder.With mixture stirring at room 4 hours under nitrogen.With mixture with ether dilution and wash with water.With the organic layer dried over mgso, filter and concentrate.With resistates flash chromatography purifying, obtain compound 271 with ethyl acetate/hexane stage gradient wash-out.

Reaction scheme 67

The available Wittig of (embodiment 5)/alkylene method as mentioned above, or adopt and to relate to the connection of on the method that C17 contains the compound of methyl ketone such as ketone 275 is containing the compound of C17 ketone functional group, finishing side chain.The compound that contains C17 ketone functional group can divide for four steps finished by the method described in the document to the transformation of methyl ketone derivative.Reaction scheme 68 has illustrated an example of this method.

Reaction scheme 68

The first step of aforesaid method comprises with acetylene lithium-ethylene diamine complex and changes compound 271 into alkynol compound 272.The acetylene lithium-complex is suspended among the THF, be cooled to-20 ℃ after, add the THF solution of compound 271.With this mixture stirring at room 6 hours, be cooled to 0 ℃ after the water termination reaction.With the mixture dichloromethane extraction, with the organic extract liquid salt solution and the water washing that merge, use dried over mgso then.After filtering and concentrating, resistates is obtained compound 272 with flash chromatography on silica gel (1: 6 ethyl acetate/hexane) purifying.

Compound 272 dehydrations are obtained the conjugated carbon-to-carbon double bond at the D of compound 273 in the ring.Compound 272 is dissolved in anhydrous pyridine and adds phosphoryl chloride.With mixture stirring at room 30 minutes, pour in the frozen water then.Aqueous slurry with dichloromethane extraction and with the organic extract liquid sodium bicarbonate aqueous solution and the water washing that merge, is used dried over mgso.After filtering and concentrating, crude product is obtained compound 273 with flash chromatography on silica gel (1: 6 ethyl acetate/hexane) purifying.

Use the Dowex-50w resin of mercury (2+) infiltration in the solvent mixture of methyl alcohol, THF and water, to prepare conjugated methyl ketone 274 then from compound 273.Compound 273 is dissolved in methyl alcohol/THF (2: 1) and adds 2 drip.Add Hg ²⁺/ Dowex resin also down stirs mixture in 60 ℃ and to spend the night.Filter and concentrate and obtain resistates, it is obtained methyl ketone 274 by flash chromatography on silica gel (1: 2 ethyl acetate/hexane) purifying.

With the Δ in the compound 274 ¹⁶Carbon-to-carbon double bond SODIUM HYDROSULPHITE sodium reduction.The toluene solution of compound 274 is joined in the aqueous solution of V-Brite B and sodium bicarbonate mixture.Add phase-transfer catalyst Aliquat ^336 and mixture refluxed 3 hours.With mixture with dichloromethane extraction and with the organic phase dried over mgso.Filter and the filtrate vacuum concentration obtained containing the crude product of compound 275.

This section or in coupling behind the suitable side chain; can be according to (tetrahedrons such as Horita; 1986; 42 (11); 3021-3028) described method is with the C15 hydroxyl of compound 275 deprotection optionally; described method comprise with to methoxy-benzyl ether with 2,3-two chloro-5,6-dicyano benzoquinone (DDQ) is in methylene dichloride and water oxygenization.Then, available several different methods, comprise with PCC and in methylene dichloride, the C15 hydroxyl oxygen that generates changed into the ketone functional group.

Embodiment 7

As described in the introductory section of this section, compound can be introduced with the Grignard method as the hemiacetal side chain functional group in 165.At first the process two-step reaction is transformed into the compound that contains C22 aldehyde with the methyl ketone functional group of C17.There are many working groups that the method (as described in reaction scheme 69) that is used for this transformation is described, comprise Koreeda etc., tetrahedron, 19,1641-1644,1978.

Reaction scheme 69

The coupling of required side chain and steroid aldehyde 278 (reaction scheme 70) is finished by the nucleophilic attack of carboanion.Compound 280 prepares (referring to the 3rd part, embodiment 9, reaction scheme 72) on the spot by handling iodide 284 down in-78 ℃ with tert-butyl lithium in ether, then to the diethyl ether solution that wherein adds aldehyde 278.This mixture was stirred 1.5 hours down in-78 ℃, be warming up to room temperature then and add ethyl acetate.With organic layer water successively, 1N hydrochloric acid and water washing, use dried over mgso then.After filtering and concentrating, crude product is obtained compound 281 with the flash chromatography on silica gel purifying.Use ordinary method, for example handle down in 60 ℃ that the side chain deprotection is obtained required side chain hemiacetal with 80% acetic acid aqueous solution.

Reaction scheme 70

The 3rd part

Be used for synthetic with the various side chain carbon skeletons of steroid nucleus link coupled

Compound 165 and listed analogue synthetic is to converge synthetic the steroid nucleus coupling of the side chain and the functionalizing of height functionalizing (that is, with).Part 2 has been described and has been used for side chain carbon skeleton and two kinds of methods of steroidal ring structure link coupled.Described below is the preparation that is used for the side chain carbon skeleton of linked reaction.

First method relates to from the synthetic 5-acetoxy-3 of primary alconol 154 (make from the L-Karvon, the L-Karvon can be from for example Aldrich Chemical Co., Milwaukee, WI obtains)-(1 '-methylethyl)-valeral (156) and relevant aldehyde.What this reaction sequence generated is the compound 165 with S-configuration.Compound with R-configuration is synthetic from the D-Karvon.Should also be noted that and to use any suitable pure protecting group to replace acetic ester, be suitable for reaction and compound 252 or relevant steroide link coupled aldehyde cpd by the alkylene type with generation.

Second method (reaction scheme 72) relates to from carboxylic acid cpd 282 synthetic compounds 284 and relevant alkyl halide.This reaction sequence also generates the product (280) with S-configuration.Equally, the enantiomorph of compound 280 is synthetic from the D-Karvon.Equally, available any suitable aldehyde protecting group replaces the ketal functional group in 280, is used for generating the used carboanion of linked reaction (making from corresponding alkyl halide by lithium/halogenide permutoid reaction).

Embodiment 8

5-acetoxy-3-(1 '-methylethyl)-valeral (156)

Intermediate 5-acetoxy-3-(1 '-methylethyl)-valeral (156) used in the described Wittig reaction of aforementioned each several part can be synthetic according to the reaction sequence shown in the reaction scheme 71.

Reaction scheme 71

According to the method in the document (the tetrahedron communication, 1984,25 (41), 4685-4688) L-Karvon (153) is transformed into compound 154.By being transformed into acetic ester the primary alconol in the compound 154 is protected.(207mg 1.10mmol) is dissolved in pyridine (2ml) and add DMAP (10mg) and diacetyl oxide (2ml) with compound 154.With the mixture stirred overnight at room temperature, dilute with ether then.Mixture is used 10% sodium bicarbonate aqueous solution and water washing successively, use dried over mgso then.With crude product with flash chromatography on silica gel (1: 4 ethyl acetate/hexane) purifying obtain compound 155 (249mg, 1.08mmol), yield 98%.

Remove the ketal protected required aldehyde 156 that obtains in the ketal 155 with 80% acetate.(150mg 0.652mmol) is suspended in 80% acetate (5ml) and with this mixture to descend to heat 2 hours in 70 ℃ with compound 155.The vacuum steaming desolventizes and resistates is added in the ether (50ml).This mixture with sodium bicarbonate aqueous solution and salt water washing, is used dried over mgso then.Filter and evaporated filtrate obtain purified compound 156 (110mg, 0.591mmol), yield 91%.

Embodiment 9

Used intermediate 280 can be synthetic according to the reaction sequence described in the reaction scheme 72 in the described linked reaction of aforementioned each several part.

Reaction scheme 72

At first use conventional literature method (tetrahedron communication, 25 (41), 4685-4688,1984) synthetic compound 282, be converted into 280 by three-step reaction then.(1.04g 5.12mmol) is dissolved in anhydrous tetracol phenixin (120ml), adds I with compound 282 ₂(2.57g) and iodobenzene diacetate ester (IDBA) (3.28g).Mixture refluxed and with 100 watts bulb irradiation 10 minutes.After being cooled to room temperature, add 5%Na ₂S ₂O ₃The aqueous solution (300ml) becomes colorless until solution, then separatory.Organic phase is washed and uses with water dried over mgso.The purified purified compound 283 (657mg, 2.30mmol, yield 45%) that obtains.

Use 2 and use tosic acid, make compound 284 from compound 283 by exchange aldehyde protecting group as catalyzer.(42mg 0.18mmol) is dissolved in benzene (5ml) and add 2 (300mg) and tosic acid (3mg) with compound 283.This mixture heating up backflow is spent the night, and cooling and vacuum-evaporation are to doing then.Resistates is obtained compound 284 (40mg, 0.14mmol, 80%) with flash chromatography on silica gel (19: 1 dichloromethane/hexane) purifying.

At last, in THF, carry out lithium-iodine permutoid reaction with tert-butyl lithium and obtain required product 280 284.Under-78 ℃, (pentane solution of 1.7M, (52.1mg is in anhydrous diethyl ether 0.160mmol) (2.0ml) solution 0.25ml) to join compound 284 with tert-butyl lithium.This solution is stirred down in-78 ℃, no longer include raw material until the GC analysis revealed and exist.This solution is directly used in the linked reaction with aldehyde 278 (embodiment 7, reaction scheme 70).

The 4th part

3,4,6,7-poly-hydroxy-22,29-epoxy-15-ketone steroid synthetic

Have 22,29-epoxy-3,4,6,7, one of contained C15 ketone and side chain hemiacetal functional group or both polyhydroxylated steroides can synthesize with the combination of the method described in first and second parts in 29-penta hydroxy group-14 β-stigmastane-15-ketone (165).Can be earlier that A and B ring is functionalized, encircle D functionalized then and the coupling side chain, perhaps adopt opposite order.In the compound that contains C15 ketone and/or C29 hemiacetal, the A/B ring can contain 2,3 or 4 hydroxyls at carbon 3,4,6 and/or 7 arbitrary combination with arbitrary combination and configuration.

Below described 22,29-epoxy-3,6,7,29-tetrahydroxy-14 β-stigmastane-15-ketone (304) and 22,29-epoxy-3,4,6,7,29-penta hydroxy group-14 β-stigmastane-15-ketone (165) and corresponding H14 α epimer synthetic.These are examples of the compound that contains C15 ketone and side chain C29 hemiacetal that obtains by the described synthetic transformation reaction of first and second parts, and the carbon 3,4,6 and 7 that this method also can be used for being created in the A/B ring system contains the compound of other hydroxylation form.

Below also comprised 22,29-epoxy-3,6,7, the synthetic route of 29-tetrahydroxy stigmastanol (306), this compound contains polyhydroxylated A/B ring system and hemiacetal side chain contained in the compound 165, but does not contain C15 ketone functional group.

Embodiment 10

22,29-epoxy-3,6,7,29-tetrahydroxy-14 β-stigmastane-15-ketone (304)

Steroidal 22,29-epoxy-3,6,7,29-tetrahydroxy-14 β-stigmastane-15-ketone (304) can synthesize according to reaction scheme 73.

Reaction scheme 73 Annotate: (ⅰ) p-TsOH, (CH ₂OH) ₂, benzene (ⅱ) TBDMSCl, imidazoles, DMF (ⅲ) CrO ₃, 3, methylene dichloride (ⅳ) NaBH ₄, CeCl ₃, THF-MeOH (ⅴ) diacetyl oxide, pyridine (ⅵ) BH ₃-THF title complex is H then ₂O ₂, OH ^-, (ⅶ) NaH, DMF, bromotoluene (ⅷ) TBAF, THF (ⅸ) PDC, methylene dichloride (ⅹ) LS-Selectride ^, THF (ⅹ ⅰ) NaH, DMF, bromotoluene (ⅹ ⅱ) acetate, water, acetone.

Reaction scheme 73 (continuing):

(ⅹ ⅲ) EtPh ₃Br, potassium tert.-butoxide, THF (ⅹ ⅳ) 156, Me ₂AlCl, methylene dichloride (ⅹ ⅴ) sodium methylate, methyl alcohol (ⅹ ⅵ) Ag ₂CO ₃/ diatomite, benzene (ⅹ ⅶ) CrO ₃, 3, DMF (ⅹ ⅷ) H ₂, Pd/C, ethyl acetate/ethanol (ⅵ ⅹ) potassium hydroxide, methyl alcohol (ⅹ ⅹ) is (CH 1. ₂OH) ₂, p-TsOH, benzene, 2.DIBAL, toluene, 3.80% acetate.

Steroidal 22,29-epoxy-3,6,7,29-tetrahydroxy-14 β-stigmastane-15-ketone (304) can be synthetic from the steroidal dehydroepiandros-sterone (247) that can buy.At first the tosic acid of spent glycol and catalytic amount is handled in the benzene that refluxes the C17 ketone of compound 247 is protected with the form of ketal.In DMF, 3 β hydroxyls are obtained compound 286 with the form protection of silyl ether with tert-butyldimethylsilyl chloride and imidazoles then.Compound 286 usefulness chromium trioxides and 3 are carried out the allylic oxidation at C7 position carbonylate (compound 287).This ketone is carried out 1 of chemo-selective with sodium borohydride and Cerium II Chloride in THF-methanol solvate system, the 2-reduction obtains vinyl carbinol 288.

Available embodiment 1 described order is introduced C6 alcohol, or with borine-THF title complex hydroboration, introduces C6 alcohol with the alkaline hydrogen peroxide oxydrolysis then.In the second approach, the vinyl carbinol in the compound 288 at first with the form protection with acetic ester of diacetyl oxide and pyridine, is dissolved in product 289 anhydrous THF and then at 0 ℃ of THF solution that adds the 1.0M borine down.This mixture was stirred 30 minutes down in 0 ℃, and stirring at room is 2.5 hours then.Drip 3N sodium hydroxide solution and 30% aqueous hydrogen peroxide solution then successively.With mixture stirring at room 16 hours, pour in the saturated nacl aqueous solution then.With these aqueous slurries with chloroform extraction and with the organic layer dried over mgso that merges.Filtration and evaporated filtrate obtain crude product, and it is obtained compound 221 by the fast silica gel chromatogram purifying.

With ortho position 6,7-glycol 221 usefulness bromotoluenes and the sodium hydride form with the benzyloxy derivative in dimethyl formamide is protected then.Compound 221 is dissolved in dimethyl formamide and adds sodium hydride.With this mixture stirring at room 1 hour, add bromotoluene then.Continue to stir 2.5 hours, add the entry termination reaction then and continue and stirred 30 minutes.With the mixture extracted with diethyl ether, then successively with 5% hydrochloric acid, saturated sodium bicarbonate solution and saturated nacl aqueous solution washing.With the organic layer dried over mgso, filter and be evaporated to dried.The crude product that obtains is obtained compound 291 with the flash chromatography purifying.

Reflux in THF with TBAF and the C3 in the compound 291 alcohol deprotection to be obtained compound 292 in 2 hours.Obtain ketone 293 with the PDC oxidation subsequently.For finishing this transformation, crude product 292 is dissolved in methylene dichloride and adds PDC.With mixture stirring at room 22 hours.Use diatomite filtration, evaporated filtrate also obtains the product ketone 293 of white solid with the flash chromatography purifying.

293 selective reductions are generated Alpha-hydroxy with 83% yield at C3.For this reason, the THF solution with compound 293 is cooled to-78 ℃ and the slow LS-Selectride of adding ^, under nitrogen, stirred 1 hour then in-78 ℃.Add methyl alcohol and reaction mixture is warming up to room temperature.After conventional aftertreatment, obtain compound 294 with about 80% yield by the flash chromatography purifying.Yield with about 10% obtains 3 β-epimer.With the form protection of the Alpha-hydroxy in the compound 294 with benzyloxy derivatives.For this reason, compound 294 is dissolved in dimethyl formamide and adds sodium hydride.With this mixture stirring at room 1 hour, add bromotoluene then.Continue to stir 16 hours, slowly add the entry termination reaction then and continue and stirred 30 minutes.With the mixture extracted with diethyl ether, then successively with 5% hydrochloric acid, saturated sodium bicarbonate solution and saturated nacl aqueous solution washing.With the organic layer dried over mgso, filter and be evaporated to dried.The crude product that obtains is obtained compound 295 with the flash chromatography purifying.

Mixture with acetate, water and acetone (2: 1: 2) refluxes 14 hours with the C17 ketal deprotection in the compound 295, and the yield with 99% behind the purifying obtains compound 296.The ethyl triphenyl phosphonium bromide handled making ylide with potassium tert.-butoxide in THF, compound 296 is transformed into alkene 297 with this ylide.In methylene dichloride, finish the coupling of side chain carbon structure with aldehyde 156 (reaction scheme 45) and Lewis acid, dimethylaluminum chloride, obtain C22 epimer 298a and 298b behind the purifying.Use then sodium methylate in methyl alcohol with C29 acetoxyl group deprotection.The silver carbonate that glycol 299 usefulness that generate are adsorbed on the diatomite is oxidized to delta-lactone in the benzene that refluxes then.Compound 299 is dissolved in benzene, adds the silver carbonate that is adsorbed on the diatomite and also this mixture was refluxed 12 hours.Then reaction mixture is filtered, evaporation also obtains lactone 300 with resistates by the flash chromatography purifying.Compound 300 usefulness chromium trioxides and 3 are carried out the allylic oxidation in methylene dichloride, at C15 carbonylate (compound 301).With hydrogen and palladium charcoal in ethyl acetate with the conjugated Δ ¹⁶The carbon-to-carbon double bond reduction obtains compound 302.Remove the benzoic ether group in the ester 302 and make the C14 epimerization obtain containing the trihydroxy-product 303 that connects along the C/D ring and containing the trihydroxy-product that anti-C/D ring connects simultaneously, these two kinds of products separate by chromatography.At last, with form the protection ((CH of C15 ketone with ethylene ketal ₂OH) ₂, p-TsOH, benzene) and under-78 ℃, become lactol and deprotection (80% acetate) to obtain 22 the delta-lactone selective reduction with DIBAL subsequently, 29-epoxy-3,6,7,29-tetrahydroxy-14 β-stigmastane-15-ketone (304).

Perhaps, can with compound 221 usefulness for example 80% acetic acid treatment to remove all protecting groups.Then, the hydroxyl in the B-ring is optionally protected.For this reason, can use 2,2-Propanal dimethyl acetal and camphorsulfonic acid.Then, can pass through Wittig reaction, for example the ketone group of C17 is transformed into exocyclic double bond, on C17, form the ethylidene substituting group with ethyl triphenyl phosphonium bromide, potassium tert.-butoxide and THF.Then, available for example oxalyl chloride, DMSO, triethylamine change into carbonyl with the C3 hydroxyl oxygen in methylene dichloride, subsequently with the C3 carbonyl LS-Selectride that forms ^(WI) reduction obtains 3 α hydroxyls for AldrichChemical Co., Milwaukee.Then the hydroxyl deprotection in the B-ring is obtained compound 330.Reaction scheme 79 has been described the alternative method of this synthetic compound 330.

Embodiment 11

Compound 306 can be synthetic from compound 300 according to the reaction sequence in the reaction scheme 74.

Reaction scheme 74

Annotate: (ⅰ) H ₂, Pd/C, ethyl acetate/ethanol (ⅱ) DIBAL, toluene ,-78 ℃.

Compound 306 can be synthetic from compound 300 with two-step approach, and the first step is with hydrogen and palladium charcoal deprotection and simultaneously with Δ in ethyl acetate and ethanol with hydroxyl ¹⁶The carbon-to-carbon double bond reduction.With said mixture stirring at room 12 days, filter and obtain 305 by the flash chromatography purifying.According to following method the delta-lactone selective reduction is become lactol then.Compound 305 is dissolved in THF and is cooled to-78 ℃, add DIBAL and also this mixture was stirred 3 hours in-78 ℃.Aftertreatment through routine also obtains 22 with the flash chromatography on silica gel purifying, 29-epoxy-3,6,7,29-tetrahydroxy stigmastanol (306).

Embodiment 12

Reaction conditions described in the aforementioned each several part can be used for the compound 165 described in the building-up reactions route 75.

Reaction scheme 75

Annotate: (ⅰ) p-TsOH, (CH ₂OH) ₂, benzene (ⅱ) CrO ₃, 3, methylene dichloride (ⅲ) NaBH ₄, CeCl ₃, THF-MeOH (ⅳ) TBDMSCl, imidazoles, DMF (ⅴ) m-CPBA, methylene dichloride (ⅵ) 80% acetate (ⅶ) TBAF, THF (ⅷ) 2,2-Propanal dimethyl acetal, CSA, DMF (ⅸ) NaBH ₄, methyl alcohol (ⅹ) TBDMSCl, imidazoles, DMF (ⅹ ⅰ) 1.Li/NH ₃-THF, 2.Cl (O) P (OEt) ₂(ⅹ ⅱ) Li/NH ₃, the trimethyl carbinol

Reaction scheme 75 (continuing):

Annotate: (ⅹ ⅲ) TBAF, THF (ⅹ ⅳ) oxalyl chloride, DMSO, triethylamine (ⅹ ⅴ) m-CPBA, methylene dichloride (ⅹ ⅵ) acetate (ⅹ ⅶ) salt of wormwood, methyl alcohol (ⅹ ⅷ) EtPPh ₃Br, potassium tert.-butoxide, THF (ⅹ ⅸ) BnBr, NaH, DMF (ⅹ ⅹ) 156, Me ₂AlCl, methylene dichloride (ⅹ ⅹ ⅰ) sodium methylate, methyl alcohol, THF (ⅹ ⅹ ⅱ) Ag ₂CO ₃/ diatomite, benzene (ⅹ ⅹ ⅲ) CrO ₃, 3, methylene dichloride.

Reaction scheme 75 (continuing):

Annotate: (ⅹ ⅹ ⅳ) H ₂, Pd/C, ethyl acetate (ⅹ ⅹ ⅴ) KOH, methyl alcohol (ⅹ ⅹ ⅵ) is (CH 1. ₂OH) ₂, pTsOH, benzene, 2.DIBAL, toluene ,-78 ℃ of 3.80% acetate.

22,29-epoxy-3,4,6,7,29-penta hydroxy group-14 β-stigmastane-15-ketone (165) can be synthetic from steroidal 4-androstene-3,17 diketone (1) that can buy.From compound 1 synthetic intermediate 128 at synthetic 3 α, 4 β, 6 α, 7 β, 17 β-penta hydroxy group-5 α-androstanol are described (embodiment 3, reaction scheme 61) when (241).

The preparation of D-ring that is used for the compound 128 of side chain coupling method described in the following embodiment (embodiment 16, reaction scheme 79) is from removing the silyl on steroidal 128 C17.Compound 128 is dissolved in THF and adds TBAF (the THF solution of 1.0M).2 hours final vacuums of this mixture heating up backflow are concentrated.Resistates is obtained alcohol 129 by the flash chromatography purifying.

In methylene dichloride, the hydroxylic moiety of alcohol 129 is transformed into ketone with oxalyl chloride.Compound 129 is dissolved in methylene dichloride and in-78 ℃ of dichloromethane solutions that add down oxalyl chlorides.Stirring is after 15 minutes down in-78 ℃, and the adding triethylamine also continues to stir 5 minutes.The compound 142 that obtains white solid through the aftertreatment and the purifying of routine.

With compound 142 being transformed into compound 145 with the identical method of compound 241 that compound 128 is transformed into described in the embodiment 3 (reaction scheme 61).Compound 142 usefulness m-CPBA are carried out epoxidation generate epoxide 143 in methylene dichloride, carry out the epoxide ring-opening reaction with acetate subsequently and generate 3,6,7-trihydroxy--4-acetoxyl group compound 144.In methyl alcohol, remove the acetate group that is connected on the C4 oxygen with salt of wormwood and obtain compound 145.

With the reaction of the same type described in the aforementioned each several part from compound 145 synthetic compounds 165.In THF, make ylide with ethyl triphenyl phosphonium bromide and potassium tert.-butoxide, compound 145 is transformed into ethylidene compound 157 with this ylide.Then four hydroxyls are obtained benzyloxy-4 compound 158 with benzyl protection.In methylene dichloride, finish the linked reaction of side chain with aldehyde 156 (reaction scheme 71) and Lewis acid as dimethylaluminum chloride, obtain compound 159.Use then sodium methylate in methyl alcohol with C29 acetoxyl group deprotection.Then the glycol that generates is oxidized to it delta-lactone 161 with being adsorbed in silver carbonate on the diatomite in benzene.Compound 161 usefulness chromium trioxides and 3 are carried out the allylic oxidation in methylene dichloride, also simultaneously benzyl is oxidized to benzoic ether group (compound 162) at the C15 carbonylate.With hydrogen and palladium charcoal in ethyl acetate and ethanol with the conjugated Δ ¹⁶The carbon-to-carbon double bond reduction obtains compound 163.(for example methanol solution of potassium hydroxide) removes the benzoic ether group in the ester 163 and makes the C14 epimerization obtain product 164 simultaneously under alkaline condition, and this product is the epimer mixture that contains the compound that compound that cis C/D ring connects is connected with trans C/D ring.At last, with form the protection ((CH of C15 ketone with ethylene ketal ₂OH) ₂, p-TsOH, benzene) and under-78 ℃, become lactol and deprotection (80% acetate) to obtain 22 the delta-lactone selective reduction with DIBAL subsequently; 29-epoxy-3; 4,6,7; 29-penta hydroxy group-14 β-stigmastane-15-ketone (165) and its C14 epimer 22; 29-epoxy-3,4,6; 7,29-penta hydroxy group-14 α-stigmastane-15-ketone.

The 5th part

Other synthetic embodiment with novel poly-hydroxy steroide of biologic activity

Except that the compound described in the aforementioned each several part, also made the related compound that has biologic activity in a large number.These compounds comprise having 3 α, 6 α, 7 beta-hydroxy forms and the compound of various functional groups is arranged at the C17 bit strip, and have 3 β, 6 α, 7 beta-hydroxy forms and the compound of various functional groups is arranged at the C17 bit strip.The method that is used to prepare these compounds has been described in following examples.

Embodiment 13

Can from midbody compound 221 preparation a large amount of 3,6, the hydroxylated compound of 7-.As described in reaction scheme 73, can be from commercially available raw material dehydroepiandrosterone (247) preparation compound 221.Specifically, in the 500ml round-bottomed flask, (20.0g 69.3mmol) is dissolved in benzene (200ml), then this flask is connected with Stark equipment with compound 247.(0.501g 2.64mmol), adds ethylene glycol (20ml) subsequently and also this mixture was refluxed 4.5 hours to add the tosic acid monohydrate.This mixture is cooled to room temperature and uses ether (200ml) dilution.With organic layer use successively saturated sodium bicarbonate solution (2 * 100ml) and saturated nacl aqueous solution (2 * 100ml) washing.With the organic layer dried over mgso, filter and be evaporated to the dried product 285 (22.8g, 68.6mmol, 99%) that obtains, this product is not purified to be directly used in next step reaction.Can be as follows with the form protection of 3 β hydroxyls in the compound 285 with silyl ether.(22.5g 67.7mmol) is dissolved in the mixture of dimethyl formamide (112.5ml) and methylene dichloride (112.5ml) with compound 285.Add successively imidazoles (11.3g, 166.0mmol) and tert-butyldimethylsilyl chloride (15.8g, 104.8mmol).With this mixture stirring at room 6 hours under argon gas, use ether (675ml) dilution then.With organic mixture use successively 5% hydrochloric acid (2 * 135ml), saturated sodium bicarbonate solution (2 * 135ml) and saturated nacl aqueous solution (2 * 135ml) washing.With the organic layer dried over mgso, filter and be evaporated to the dried crude product 286 that obtains.This crude product is obtained white crystals shape compound 286 (total recovery in two steps is 83% for 25.9g, 58.0mmol) with ethyl acetate/methanol (3: 2) recrystallization.Then compound 286 is carried out the allylic oxidizing reaction at C7 position carbonylate (compound 287).(15.0g 33.6mmol) is dissolved in hexanaphthene (60ml) and add entry (7.3ml) with compound 286.(0.0561g 0.27mmol), drips tertbutyl peroxide (37.6ml) subsequently to add hydration ruthenium chloride (III).With this mixture stirring at room 24 hours, use ethyl acetate (376ml) dilution then.With organic mixture use successively saturated nacl aqueous solution (2 * 188ml) and 25% hypo solution (2 * 188ml) washing.With the organic layer dried over mgso, filter and be evaporated to the dried crude product that obtains.Crude product is obtained compound 287 (8.6g, 18.7mmol, 56%) with re-crystallizing in ethyl acetate.Ketone in the compound 287 is carried out 1 of chemo-selective, and the 2-reduction can obtain 288.For this reason, (14.7g 31.9mmol) is dissolved in tetrahydrofuran (THF) (118ml) and add seven hydration Cerium II Chlorides (III) (17.6g, methyl alcohol 47.2mmol) (35ml) solution with compound 287.With mixture with ice bath cooling and slowly add sodium borohydride (2.5g, 66.1mmol).This mixture is warming up to room temperature and stirred 2.5 hours.In mixture, slowly add 5% hydrochloric acid (44ml), add ethyl acetate (588ml) then.Reaction solution is used 5% hydrochloric acid (120ml), saturated sodium bicarbonate solution (120ml) and saturated nacl aqueous solution (120ml) washing successively.With the organic layer dried over mgso, filter and be evaporated to the dried product 288 (14.8g) that obtains, this product is not purified to be directly used in next step reaction.Compound 288 (14.8g) is dissolved in pyridine (30ml) and adds the 4-Dimethylamino pyridine (30mg) of diacetyl oxide (15ml) and catalytic amount.With this mixture stirring at room 16 hours, use ethyl acetate (300ml) dilution then.(saturated nacl aqueous solution (2 * 60ml) washings are used in 2 * 60ml) washings then with saturated sodium bicarbonate solution with organic mixture.With the organic layer dried over mgso, filter and be evaporated to the dried crude product that obtains.Obtain compound 289 (total recovery in two steps is 78% for 12.6g, 25.0mmol) with recrystallizing methanol.Compound 289 hydroborations are obtained 6 α, 7 β hydroxylation forms.Therefore, (8.4g 16.6mmol) is dissolved in anhydrous THF (50ml) and also this mixture is cooled to 0 ℃ with compound 289.Add THF (20ml) solution of 1.0M borine and this mixture was stirred 30 minutes in 0 ℃, stirring at room is 2.5 hours then.Drip 10N sodium hydroxide solution (10ml) and 30% aqueous hydrogen peroxide solution (10ml) then successively.With mixture stirring at room 18 hours, pour into then in the saturated nacl aqueous solution (200ml).With this aqueous slurry with methylene dichloride (2 * 250ml) extractions and with the organic layer that merges (2 * 250ml) wash, then with the organic layer dried over mgso with 25% sodium thiosulfate solution.Filtration and evaporated filtrate obtain crude product, and it is obtained compound 221 (5.9g, 12.3mmol, 74%) by flash chromatography on silica gel (3: 1 hexane/ethyl acetate) purifying.

Reaction scheme 76 has illustrated from compound 221 synthetic compounds 326 and 327.(1.2g 2.4mmol) is dissolved in diacetyl oxide (3ml) and add the 4-Dimethylamino pyridine (40mg) of pyridine (3ml) and catalytic amount with compound 221.With this mixture stirring at room 3 hours, use ethyl acetate (100ml) dilution then.Organic mixture is used 5% hydrochloric acid, saturated sodium bicarbonate solution (100ml) and saturated nacl aqueous solution (100ml) washing successively.With the organic layer dried over mgso, filter and be evaporated to the dried product 321 (1.3g) that obtains, this product is not purified to be directly used in next step reaction.In THF, remove the compound 322 that the silyl protecting group obtains containing 3 beta-hydroxies with TBAF.Crude product 321 is dissolved in THF (10ml) and adds the tetrabutyl ammonium fluoride (4ml) of 1.0M.This mixture was refluxed 1 hour, be cooled to room temperature, pour into then in the saturated nacl aqueous solution (50ml).With this aqueous slurry with methylene dichloride (5 * 40ml) extraction and with the organic layer dried over mgso.Filtration and evaporated filtrate obtain crude product, and it is obtained compound 322 (total recoverys in two steps are 76% for 0.85g, 1.9mmol) by flash chromatography on silica gel (1: 1 hexane/ethyl acetate) purifying.In methylene dichloride, carry out oxidizing reaction with PDC then and make the stereochemistry of C3 that living ketone 323 take place to transform, pass through L-Selectride subsequently ^Reduce, reduction reaction mainly generates 3 'alpha '-hydroxylation compounds 324.For this reason, with compound 322 (0.84g, 1.9mmol) be dissolved in methylene dichloride (15ml) and add PDC (1.2g, 3.2mmol).With mixture stirring at room 40 hours, use ether (50ml) dilution then.Filtration also is evaporated to the dried crude product that obtains, and it is obtained compound 323 (0.81g, 1.8mmol, 95%) by flash chromatography on silica gel (9: 1 hexane/ethyl acetate) purifying.(0.34g 0.75mmol) is dissolved in THF (10ml) and be cooled to-78 ℃ with compound 323 then.(the THF solution of 1.0M 1.6ml) and with this mixture stirred 1 hour down in-78 ℃ to add L-Selectride.Mixture is warming up to room temperature and adds 10N sodium hydroxide solution (1ml), drip 30% aqueous hydrogen peroxide solution (1ml) then.With mixture stirring at room 1 hour, pour into then in the ethyl acetate (50ml).With organic mixture use successively 5% hydrochloric acid (2 * 25ml), saturated sodium bicarbonate solution (2 * 25ml) and saturated nacl aqueous solution (2 * 25ml) washing.With the organic layer dried over mgso, filter and be evaporated to dried, obtain compound 324 (0.214g, 0.48mmol, 64%) with flash chromatography on silica gel (3: 1 hexane/ethyl acetate) purifying.

Remove the acetic ester protecting group then.(0.25g 0.56mmol) is dissolved in methyl alcohol (10ml) and add sodium methylate (250mg) with compound 324.With mixture stirring at room 3 hours, use ethyl acetate (50ml) dilution then.With organic mixture use successively 5% hydrochloric acid (2 * 25ml), saturated sodium bicarbonate solution (2 * 25ml) and saturated nacl aqueous solution (2 * 25ml) washing.With the organic layer dried over mgso, filter and be evaporated to dried, obtain compound 325 (0.185g, 0.51mmol, 91%) with flash chromatography on silica gel (ethyl acetate) purifying.(141mg 0.385mmol) is dissolved in 80% acetate (10ml) and stirred 14 hours down in 70 ℃ with compound 325.With mixture with ethyl acetate (50ml) dilution, use successively then saturated sodium bicarbonate solution (2 * 25ml) and saturated nacl aqueous solution (2 * 25ml) wash.With the organic layer dried over mgso, filter and be evaporated to dried, obtain compound 326 (0.054g, 0.17mmol, 44%) with flash chromatography on silica gel (ethyl acetate) purifying.At last, (0.023g 0.072mmol) uses sodium borohydride (0.034g) to reduce under room temperature in 95% ethanol (1ml) and generates tetrahydroxy compound 327 (0.018g, 0.056mmol, 78%) with ketone 326.

Reaction scheme 76

Annotate: (ⅰ) diacetyl oxide, pyridine (ⅱ) TBFA, THF (ⅲ) PDC, methylene dichloride (ⅳ) L-Selectride ^, THF (ⅴ) sodium methylate, methyl alcohol (ⅵ) 80% acetate (ⅶ) sodium borohydride, ethanol.

Embodiment 14

As described in reaction scheme 77, compound 329 can be synthetic with two-step approach from compound 322.Ester 322 stirring at room in sodium methylate and methyl alcohol was removed the acetic ester protecting group in 15 hours.Make compound 328 slough ketal with 80% acetate then and obtain trihydroxy-compound 329.

Reaction scheme 77

Annotate: (ⅰ) sodium methylate, methyl alcohol (ⅱ) 80% acetate.

Embodiment 15

Shown in reaction scheme 78, compound 329 can also be with 80% acetate from compound 221 directly with one-step synthesis.For this reason, (1.3g 2.7mmol) is dissolved in 80% acetic acid aqueous solution (20ml) and with this mixture stirring at room 3 hours with ketal 221.Be evaporated to the dried compound 329 (0.79g, 2.5mmol, 93%) that obtains, the not purified subsequent reaction that is directly used in of this product.

Reaction scheme 78

Embodiment 16

Steroidal 3 α, 6 α, 7 β-trihydroxy--17 (20)-pregnene (330) can be synthetic according to the reaction sequence shown in the reaction scheme 79.

Reaction scheme 79

Annotate: (ⅰ) p-TsOH, (CH ₂OH) ₂, benzene (ⅱ) CrO ₃, 3, methylene dichloride (ⅲ) NaBH ₄, CeCl ₃, THF-MeOH (ⅳ) TBDMSCl, imidazoles, DMF (ⅴ) m-CPBA, methylene dichloride (ⅵ) 80% acetate (ⅶ) TBAF, THF (ⅷ) 2,2-Propanal dimethyl acetal, CSA, DMF (ⅸ) NaBH ₄, methyl alcohol (ⅹ) TBDMSCl, imidazoles, DMF (ⅹ ⅰ) 1.Li/NH ₃, Et ₂O, 2.PDC, methylene dichloride (ⅹ ⅱ) L-Selectride ^, THF.

Reaction scheme 79 (continuing):

Annotate: (ⅹ ⅲ) diacetyl oxide, pyridine (ⅹ ⅳ) TBAF, THF (ⅹ ⅴ) is (COCl) ₂, triethylamine, DMSO (ⅹ ⅵ) EtPh ₃PBr, potassium tert.-butoxide, THF (ⅹ ⅶ) 80% acetate.

Compound 330 can be from compound 10 synthetic (shown in reaction scheme 79).Under-78 ℃ and the argon gas, (0.82g, ether 1.73mmol) (15ml) solution transfer in the flask of liquefied ammonia (30ml) solution that contains metallic lithium (55mg) with compound 10.-78 ℃ after following 30 minutes, add ammonium chloride (2.0g) and ammonia is evaporated.Add entry (10ml) and separatory.(2 * 25ml) extractions, (2 * 25ml) wash and use dried over mgso with the organic layer water that merges with methylene dichloride with water layer.Filter and be evaporated to do after, resistates is dissolved in methylene dichloride (15ml) and add PDC (600mg, 1.59mmol).With mixture stirring at room 18 hours, filter with Celite pad then.Filtrate is obtained compound 13 (653mg, 1.40mmol, 81%) by flash chromatography on silica gel (5: 1 hexane/ethyl acetate) purifying.

Then ketone 13 is reduced by the following method.(1.2g 2.53mmol) is dissolved in THF (30ml) and also this mixture is cooled to-78 ℃, adds L-Selectride with compound 13 ^(the THF solution of 1.0M, 3.8ml).The stirring under-78 ℃ of this mixture was warming up to 0 ℃ in 2.5 hours then.Add 10% sodium hydroxide solution (10ml) and 30% hydrogen peroxide (10ml) successively.Stir after 2 hours, also (4 * 100ml) extract with methylene dichloride with this aqueous slurry to add entry (20ml).With organic extract liquid 1 0%Na that merges ₂S ₂O ₃(2 * 100ml) and saturated nacl aqueous solution (2 * 100ml) washing.With the organic layer dried over mgso, filter and be evaporated to the dried product 14 that obtains, this product is not purified to be directly used in next step reaction.Then 3 Alpha-hydroxies are obtained acetic ester 15 with diacetyl oxide and pyridine with the form protection of acetic ester.For this reason, compound 14 being dissolved in pyridine (15ml) and diacetyl oxide (10ml) also stirs this mixture 12 hours.Add ethyl acetate and ether (1: 1,150ml) and with this mixture use successively 5% hydrochloric acid (2 * 50ml) and saturated sodium bicarbonate solution (2 * 50ml) wash.With the organic layer dried over mgso, filter and be evaporated to dried, with resistates by flash chromatography on silica gel (10: 1 hexane/ethyl acetate) purifying obtain compound 15 (0.89g, 1.73mmol, two the step total recoverys be 68%).By (the THF solution of 1.0M refluxes in 3.6ml) and was evaporated to dried compound 15 (0.85g, 1.63mmol) the silyl protecting group of C17 of removing in 3 hours then at THF (30ml) and TBAF.Then resistates is dissolved in (3 * 30ml) washings of methylene dichloride (100ml) and water.With the organic layer dried over mgso, filter and be evaporated to dried, resistates is obtained compound 16 (0.59g, 1.45mmol, 89%) by flash chromatography on silica gel (1: 1 hexane/ethyl acetate) purifying.Use oxalyl chloride C17 hydroxyl oxidize with compound 16 in DMSO then.For this reason, with compound 16 (0.57g, 1.40mmol) be dissolved in methylene dichloride (5ml) and under-78 ℃, join oxalyl chloride (0.15ml, 1.68mmol) and DMSO (0.24ml is in methylene dichloride 3.36mmol) (10ml) solution.-78 ℃ were stirred down after 15 minutes, added triethylamine (0.98ml) and continued and stirred 5 minutes.Mixture is warming up to room temperature and adds entry (10ml).Separatory and with organic layer use successively 5% hydrochloric acid (2 * 5ml) and saturated sodium bicarbonate solution (2 * 5ml) washing.With the organic layer dried over mgso, filter and be evaporated to dried, resistates is obtained compound 17 (0.54g, 1.33mmol, 95%) by flash chromatography on silica gel (2: 1 hexane/ethyl acetate) purifying.

In THF, make phosphorus ylide with ethyl triphenyl phosphonium bromide and potassium tert.-butoxide, with this ylide compound 17 is carried out Wittig reaction and generate compound 18.Under the nitrogen gas stream, (0.59g, (1.94g is in THF 5.23mmol) (15ml) solution and with this mixture stirring at room 1 hour 5.23mmol) to join the ethyl triphenyl phosphonium bromide with potassium tert.-butoxide.(0.53g 1.31mmol) is dissolved in THF (10ml) and also this solution is joined in the THF solution of ylide with compound 17.The mixture that forms refluxed under nitrogen 12 hours, is cooled to room temperature then.With reaction mixture with diatomite filtration and filtrate is evaporated to dried.With resistates be dissolved in methylene dichloride (10ml) and with saturated ammonium chloride solution (2 * 30ml) and water (2 * 30ml) wash.With the organic layer dried over mgso, filter and be evaporated to dried, resistates is obtained compound 18 (0.33g, 0.88mmol, 67%) by flash chromatography on silica gel (3: 1 hexane/ethyl acetate) purifying.

At last, remove the acetone solvate group.For this reason, (20mg 0.053mmol) is dissolved in 80% acetic acid aqueous solution (1.5ml) and stirred 1 hour in 60 ℃ with compound 18.Mixture is evaporated to the dried compound 330 (17.8mg, 0.053mmol, 99%) that obtains.

Embodiment 17

Can have important biomolecule in a large number and learn active compound from compound 329 is synthetic.For example, contain 3 β, 6 α, 7 beta-hydroxy forms and can make (reaction scheme 80) by three-step reaction from compound 329 at the compound 333 that C17 has an ethylidene.With compound 329 (1.81g 5.6mmol) is dissolved in 2,2-Propanal dimethyl acetal (25ml) and the camphorsulfonic acid (CSA) that adds catalytic amount (0.03g), then with this mixture stirring at room 3 hours.Add ethyl acetate (200ml) and with this mixture use successively 5% hydrochloric acid (50ml), saturated sodium bicarbonate solution (2 * 100ml) and saturated nacl aqueous solution (2 * 100ml) wash.With the organic layer dried over mgso, filter and be evaporated to dried, resistates is obtained compound 331 (1.54g, 4.3mmol, 76%) by flash chromatography on silica gel (1: 1 hexane/ethyl acetate) purifying.Carry out Wittig reaction with phosphorus ylide described in the aforementioned each several part and compound 331 and generate compound 332.Under the nitrogen gas stream, (7.15g, (23.7g is in toluene 63.7mmol) (360ml) solution 63.7mmol) to join ethyl triphenyl phosphonium bromide in the stirring with potassium tert.-butoxide.With this mixture stirring at room 1 hour, add compound 331 (7.7g, toluene 21.2mmol) (210ml) solution then.With this mixture stirring at room 24 hours under nitrogen, drip water (120ml) termination reaction then.With mixture with ethyl acetate (900ml) dilution, use then saturated sodium bicarbonate solution (2 * 200ml) and sodium chloride solution (2 * 200ml) wash.With the organic layer dried over mgso, filter and be evaporated to dried, resistates is obtained compound 332 (7.2g, 19.2mmol, 90%) by flash chromatography on silica gel (2: 1 hexane/ethyl acetate) purifying.Then by the hydroxyl deprotection in 80% acetate, stirring compound 332 with 332.For this reason, (7.2g 19.2mmol) is dissolved in 80% acetate (115ml) and with this mixture stirring at room 3 hours with compound 332.Mixture is evaporated to dried, obtains compound 333 (5.81g, 17.4mmol, 90%) by flash chromatography on silica gel (9: 1 methylene chloride) purifying subsequently.

Reaction scheme 80

Embodiment 18

Contain ketone and have 6 at C3, the compound of 7-hydroxylation form can make from compound 332.For example, compound 332 oxidation under the Swern condition is generated compound 334, then its deprotection is generated compound 335 (reaction scheme 81).(1.01g 2.70mmol) is dissolved in methylene dichloride (10ml), then in-78 ℃ of dichloromethane solutions (8.1ml) that add DMSO (2.5ml) and 2.0M oxalyl chloride with compound 332., add triethylamine (4.6ml) and continue and stirred 15 minutes after 15 minutes in-78 ℃ of stirrings, stirring at room is 30 minutes then.With mixture with ethyl acetate (100ml) dilution and with saturated sodium bicarbonate solution (2 * 50ml) and saturated nacl aqueous solution (2 * 50ml) wash.With the organic layer dried over mgso, filter and be evaporated to dried, resistates is obtained compound 334 (0.77g, 2.07mmol, 77%) by silica gel (with the hexane solution pre-treatment of 1% triethylamine) flash chromatography (19: 1 hexane/ethyl acetate) purifying.According to the description in the previous embodiment; with compound 334 (11mg; 0.030mmol) in 80% acetate (1.25ml) stirring at room 1 hour with the hydroxyl deprotection in the compound 334; be evaporated to dry doubling and obtain compound 335 (9.8mg after by flash chromatography on silica gel (ethyl acetate) purifying; 0.029mmol, 97%).

Reaction scheme 81

Embodiment 19

The Swern oxidizing reaction by product is a chlorinated derivative 336 in the reaction scheme 81.Shown in reaction scheme 82, can be by handling with compound 336 deprotections with 80% acetic acid aqueous solution.For this reason, (0.028g 0.072mmol) is dissolved in 80% acetic acid aqueous solution (2ml) and stirring at room 1 hour with compound 336.Mixture is evaporated to dried, resistates obtains compound 337 (0.024g, 0.067mmol, 94%) by flash chromatography on silica gel (3: 2 hexane/ethyl acetate) purifying.

Reaction scheme 82

Embodiment 20

As previously mentioned, compound 330 can be by (for example, using LS-Selectride with the C3 carbonyl reduction in the compound 334 ^, Aldrich Chemical Co., Milwaukee, WI) obtain 3 α hydroxyls and subsequently deprotection make.For this reason, (0.85g 2.3mmol) is dissolved in THF (25ml) and be cooled to-78 ℃ with compound 334.(the THF solution of 1.0M 3.0ml) and with mixture stirred 3 hours in-78 ℃ to add LS-Selectride.Add 10N aqueous sodium hydroxide solution (2ml) and 30% hydrogen peroxide (2ml) and this mixture is warming up to 0 ℃.With mixture with ethyl acetate (150ml) dilution, use then saturated sodium bicarbonate solution (2 * 50ml) and saturated nacl aqueous solution (2 * 50ml) wash.With the organic layer dried over mgso, filter and be evaporated to dried, resistates obtains compound 338 (0.703g, 1.88mmol, 80%) by flash chromatography on silica gel (3: 1 hexane/ethyl acetate) purifying.Then the hydroxyl deprotection on the B-ring is obtained compound 330 (reaction scheme 83).(1.44g 3.85mmol) is dissolved in 80% acetic acid aqueous solution (25ml) and stirring at room 3 hours with compound 338.Mixture is evaporated to the dried compound 330 (1.25g, 3.74mmol, 97%) that obtains.This method is the alternative synthetic route of the synthetic compound 330 shown in the reaction scheme 79.

Reaction scheme 83

Embodiment 21

The compound that contains ethyl or other alkyl chain at C17 can be from containing the respective compound preparation of exocyclic double bond at C17.For example, compound 339 by with the two key catalytic hydrogenations of the C17-C20 in the compound 338 and subsequently deprotection make (as follows).For this reason, (0.15g 0.40mmol) is dissolved in 1: 1 acetate and ethanol (4ml) and add 10% palladium charcoal (15mg) with compound 338.Mixture was stirred 16 hours under nitrogen atmosphere, filter and be evaporated to the dried required product 339 (0.115g, 0.340mmol, 85%) (reaction scheme 84a) that obtains then.If hydrogenation carries out, can form intermediate 17-ethyl-6,7-acetone solvate 361 (reaction scheme 84b) in ethyl acetate.For this reason, (0.019g 0.050mmol) is dissolved in ethyl acetate (5ml) and add 10% palladium charcoal (9mg) with compound 338.Mixture was stirred 14 hours under nitrogen atmosphere, filter and be evaporated to the dried required product 361 (0.018g, 0.048mmol, 96%) that obtains then.

Equally, the acetic acid solution with compound 332 can obtain product 340 (reaction scheme 85) with hydrogen and 10% palladium hydrogenated carbon.

Reaction scheme 84a

Reaction scheme 84b

Reaction scheme 85

The compound that embodiment 22C17 has other alkyl can make with different Witting reagents.For example, compound 342 be easy to by with aforementioned similar Wittig reaction, use MePh ₃PBr replaces EtPh ₃PBr is as Witting reagent and subsequently triol 341 deprotections are made (reaction scheme 86).For this reason, with first base three phenyl phosphonium bromides (1.97g, 5.51mmol) and potassium tert.-butoxide (0.61g 5.4mmol) stirred in THF (10ml) 1 hour.(0.411g, THF 1.14mmol) (5ml) solution also refluxes this mixture 3 hours to add ketone 331 in this ylide.After carrying out conventional aftertreatment according to above description, crude product is obtained compound 341 (0.332g, 0.920mmol, 81%) by flash chromatography on silica gel (3: 1 hexane/ethyl acetate) purifying.(0.204g 0.567mmol) handled 1.5 hours under room temperature with 80% acetate (4ml) with compound 341 then.Mixture vacuum-evaporation is obtained required triol 342 (0.173g, 0.540mmol, 95%).Then with compound 341 (0.023g; 0.063mmol) with 10% palladium charcoal (15mg) hydrogenation under nitrogen atmosphere in ethyl acetate (5ml); subsequent filtration also is evaporated to dried; prepare corresponding C 17 methyl-derivatives with 80% acetic acid solution (2.5ml) deprotection; make compound 343 (0.018g thus; 0.056mmol, 88%) and (reaction scheme 87).

Reaction scheme 86

Reaction scheme 87

Embodiment 23

Containing the methylene radical carbon compound at C3 can be synthetic by multiple diverse ways.Wherein a kind of method comprises the improved Barton method shown in the reaction scheme 88 (Robins etc., JACS (J.Am.Chem.Soc.), 105:4059-4065,1983).(0.10g, 0.27mmol) (0.45ml 3.3mmol) handled 2 hours under room temperature in pyridine (2ml) and methylene dichloride (3ml) with chloro thion formic acid (chlorothionoformate) phenyl ester with alcohol 332.Mixture is evaporated to dry doubling resistates is obtained thion acid esters (thionester) 344 (0.12g, 0.22mmol, 84%) by flash chromatography on silica gel (30: 1 hexane/ethyl acetate) purifying.Under atmosphere of inert gases, (0.091g 0.17mmol) uses nBu with ester 344 then ₃(60 μ l, 0.22mmol) AIBN (4mg) with catalytic amount handled 3 hours down in 75 ℃ in toluene (3ml) SnH.Mixture is evaporated to dried, with obtaining compound 345 (0.035g, 0.1mmol, 57%) behind flash chromatography on silica gel (30: the 1 hexane/ethyl acetate) purifying.(0.025g 0.069mmol) with 80% acetic acid aqueous solution (2ml) room temperature treatment 1 hour, is evaporated to dry doubling then and obtains glycol 346 (0.021g, 0.066mmol, 96%) with flash chromatography on silica gel (3: 1 hexane/ethyl acetate) purifying with compound 345.

Reaction scheme 88

Embodiment 24

Containing the methylene radical carbon compound at C17 can be according to making to reaction similar described in the embodiment 23.For example; at first by using tert-butyldimethylsilyl chloride (0.095g; 0.63mmol) and imidazoles (0.057g; 0.84mmol) in dimethyl formamide (4ml), under room temperature, handled 4 hours; (0.15g, 0.42mmol) alcohol groups in is with the form protection of silyl ether with compound 331.With this mixture with ether (75ml) dilution and with saturated sodium bicarbonate solution (2 * 25ml) and water (2 * 25ml) wash.With the organic layer dried over mgso, filter and be evaporated to dried, resistates is obtained compound 347 (0.18g, 0.38mmol, 90%) (reaction scheme 89) by flash chromatography on silica gel (9: 1 hexane/ethyl acetate) purifying.Then the ketone functional group of compound 347 is reduced in ether with lithium aluminium hydride and obtain 17 β-alcohol 348.For this reason, with compound 347 (0.18g 0.37mmol) is dissolved in ether (5ml), be cooled to 0 ℃ and add lithium aluminium hydride (0.018g, 0.48mmol).Mixture in 0 ℃ of stirring 30 minutes, is dripped saturated sodium bicarbonate solution (1ml) then.With mixture with ether (50ml) dilution, with saturated sodium bicarbonate solution (2 * 15ml) and water (2 * 15ml) washings.With the organic layer dried over mgso, filter and be evaporated to dried, resistates is obtained compound 348 (0.15g, 0.31mmol, 85%) by flash chromatography on silica gel (4: 1 hexane/ethyl acetate) purifying.Adopt the Barton reaction, with compound 348 usefulness sodium hydrides, CS ₂Handle in THF with methyl iodide, behind aftertreatment and purifying, obtain methyl xanthate 349.For this reason, with compound 348 (0.052g, 0.11mmol) be dissolved in THF (5ml) and add sodium hydride (17.4mg (60% oil suspension), 0.43mmol) and imidazoles (5mg, 0.074mmol).With this mixture stirring at room 30 minutes, add dithiocarbonic anhydride (0.2ml) then and continue and stirred 2 hours, refluxed then 30 minutes.Add methyl iodide (0.2ml) and continue and refluxed 30 minutes.Drip water (1ml), with this mixture with ether (100ml) dilution, use successively then 5% hydrochloric acid (2 * 30ml), saturated sodium bicarbonate solution (2 * 30ml) and water (2 * 30ml) wash.With the organic layer dried over mgso, filter and be evaporated to dried, resistates obtains compound 349 (0.054g, 0.09mmol, 85%) after with flash chromatography on silica gel (15: 1 hexane/ethyl acetate) purifying.In next step reaction, use AIBN usually as radical initiator.For this reason, (0.05g 0.087mmol) is dissolved in toluene (15ml), adds nBu with compound 349 ₃(0.051g, 0.17mmol) AIBN's SnH (10mg) with catalytic amount also refluxes this mixture 22 hours under atmosphere of inert gases.Mixture is cooled to room temperature, is evaporated to dry doubling and obtains compound 350 (0.010g, 0.022mmol, 25%) after by flash chromatography on silica gel (40: 1 hexane/ethyl acetate) purifying.(0.010g 0.022mmol) with 80% acetate (2ml) room temperature treatment 18 hours, is evaporated to dry doubling then and obtains compound 351 (0.0065g, 0.021mmol, 96%) with flash chromatography on silica gel (20: 1 chloroform/methanol) purifying with compound 350.

Reaction scheme 89

Embodiment 25

Having the senior alkyl chain to be connected compound on the C17 can adopt with similar methods described in the above embodiment and make.For example, shown in reaction scheme 90, compound 354 can make from the cholesteryl acetate (228) that can buy by four-step reaction.Described method comprises the RuCl with C7 ₃With the tBUOOH oxidation, use NaBH then ₄/ CeCl ₃The reduction of C7 ketone is obtained alcohol 352.With compound 352 acetylizes and subsequently hydroboration (handling with alkaline peroxide subsequently) obtain required compound 354.

Specifically, with compound 228 (0.431g, 1.01mmol), RuCl ₃(0.020g, 0.098mmol), the aqueous solution of hexanaphthene (5ml), water (0.25ml) and 70%tBuOOH (1.5ml, 11.0mmol) stirring at room 24 hours together.Mixture with ethyl acetate (125ml) dilution, is used 10%Na then successively ₂SO ₃The aqueous solution (2 * 50ml) and saturated nacl aqueous solution (2 * 50ml) washing.With organic layer with dried over mgso and be evaporated to dried.Obtain compound 229 (0.263g, 0.591mmol, 59%) by flash chromatography on silica gel (9: 1 hexane/ethyl acetate) purifying.The reduction of carrying out C7 ketone as described below.With CeCl ₃7H ₂O (2.00g, 5.368mmol) and the mixture of methyl alcohol (5ml) join ketone 229 (1.11g be in THF 2.52mmol) (5ml) solution and this mixture is cooled to 0 ℃.(0.119g 5.14mmol) and with this mixture stirred 1 hour in 0 ℃, was warming up to room temperature then and continued and stirred 2 hours to add sodium borohydride.In mixture, add 5% hydrochloric acid (10ml) termination reaction carefully, use ethyl acetate (250ml) dilution then.With this milk sap use successively 5% hydrochloric acid (2 * 100ml), saturated sodium bicarbonate solution (2 * 100ml) and saturated nacl aqueous solution (2 * 100ml) washing.With organic phase with dried over mgso and be evaporated to dried.Resistates is obtained alcohol 352 (0.850g, 1.91mmol, 76%) by flash chromatography on silica gel (9: 1 hexane/ethyl acetate) purifying.Protect then and hydroboration.For this reason, with compound 352 (0.850g, 1.91mmol), the mixture stirring at room of pyridine (5ml) and diacetyl oxide (5ml) 16 hours.With this mixture with ethyl acetate (150ml) dilution, use successively then 5% hydrochloric acid (3 * 50ml), saturated sodium bicarbonate aqueous solution (2 * 50ml) and saturated sodium-chloride water solution (2 * 50ml) wash.With organic phase with dried over mgso and be evaporated to dried.Resistates is obtained product 353 (0.823g, 1.70mmol, 99%) by flash chromatography on silica gel (19: 1 hexane/ethyl acetate) purifying.(0.275g, THF 0.5648mmol) (5ml) solution are cooled to 0 ℃ and add BH with diacetate esters 353 ₃THF solution (1.0M, 2.5ml, 2.5mmol).Mixture in 0 ℃ of stirring 3 hours, is added 10N aqueous sodium hydroxide solution (1ml) and 30% aqueous hydrogen peroxide solution (1ml) termination reaction then carefully.The mixture that forms was stirred 16 hours,, use 10%Na then with ethyl acetate (100ml) dilution ₂SO ₃The aqueous solution (2 * 50ml), saturated sodium bicarbonate aqueous solution (2 * 50ml) and saturated sodium-chloride water solution (2 * 50ml) washing.With organic phase with dried over mgso and be evaporated to dried.Obtain product 3 β-acetoxyl group-6 α by flash chromatography on silica gel (3: 1 hexane/ethyl acetate) purifying; 7 beta-dihydroxyies-5 α-cholestane (0.032g; 0.069mmol; 13%); with its with sodium methylate (from sodium Metal 99.5 (0.262g, 11.4mmol) and methyl alcohol (10ml) make) under room temperature the processing 1.5 hours deprotections.With mixture with ethyl acetate (30ml) dilution, use then saturated sodium bicarbonate aqueous solution (2 * 15ml) and saturated sodium-chloride water solution (2 * 15ml) wash.With organic phase with dried over mgso and be evaporated to dried.Resistates is obtained triol 354 (0.029g, 0.069mmol, 99%) by flash chromatography on silica gel (1: 1 hexane/ethyl acetate) purifying.

Reaction scheme 90

Embodiment 26

Also prepared in the A-ring and contained the compound of other functional group and tested its biologic activity.Shown in reaction scheme 91, can make compound 360 from compound 335 by multistep is synthetic.Compound 335 usefulness diacetyl oxides and pyridine and DMAP acetylize are obtained diacetoxy compound 355.With compound 335 (1.5g, 4.5mmol), pyridine (10ml), diacetyl oxide (5ml) and 4-Dimethylamino pyridine (0.028g, mixture stirring at room 0.23mmol) 12 hours.With mixture with ethyl acetate (300ml) dilution, use successively then 5% hydrochloric acid (3 * 50ml), saturated sodium bicarbonate aqueous solution (3 * 50ml) and water (3 * 50ml) wash.With dried over mgso and be evaporated to driedly, the crude product 355 (1.9g) that obtains is not purified to be directly used in next step reaction with organic phase.(0.800g 1.9mmol) is dissolved in acetate and add 10% palladium charcoal (80mg) with crude product 355.With this mixture stirring at room 16 hours under nitrogen atmosphere.Mixture filtered and evaporation as for obtaining crude product, it is obtained compound 356 (0.702g, 1.67mmol, 88%) by flash chromatography on silica gel (5: 1 hexane/ethyl acetate) purifying.Then with compound 356 and HONH ₂-HCl refluxes in methyl alcohol-pyridine solution together and makes oxime 357.Therefore, (0.05g 0.12mmol) is dissolved in the mixture of pyridine (2ml) and methyl alcohol (2ml) and add HONH with compound 356 ₂-HCl (0.017g, 0.24mmol).Mixture refluxed 1.5 hours and with ethyl acetate (50ml) dilution, use successively then 5% hydrochloric acid (3 * 15ml), saturated sodium bicarbonate aqueous solution (3 * 15ml) and water (3 * 15ml) wash.With the organic phase dried over mgso, filter and be evaporated to the dried compound 357 (0.052g, 0.12mmol, 99%) that obtains, this product is not purified to be directly used in next step reaction.With product 357 (0.071g 0.16mmol) is dissolved in pyridine (13mg) and diacetyl oxide (3ml), be cooled to 0 ℃ and add Acetyl Chloride 98Min. (15.5mg, 0.20mmol).Then mixture was heated 8 hours in 100 ℃.Add entry (0.5ml) and continue heating 30 minutes.Mixture is cooled to room temperature, and water (10ml) dilutes and (3 * 15ml) extract with methylene dichloride.With organic extract liquid water (2 * 10ml) washings that merge.With the organic layer dried over mgso, filter and be evaporated to dried, resistates obtains compound 358 (0.41g, 0.086mmol, 52%) by flash chromatography on silica gel (2: 1 hexane/ethyl acetate) purifying.(0.020g 0.042mmol) reduces C3 ketone with sodium borohydride (2.4mg) room temperature treatment 1 hour in THF (2ml) with compound 358 then.Add acetate (2) and with this mixture with ethyl acetate (50ml) dilution, use successively then saturated sodium bicarbonate solution (2 * 15ml) and water (2 * 50ml) wash.With the organic phase dried over mgso, filter and be evaporated to dried, then crude product 359 is dissolved in methyl alcohol (1.5ml).Add sodium methylate (10mg) and with mixture stirring at room 48 hours.Add Amberlite IR-120 ion exchange resin to pH6.Mixture is filtered and is evaporated to dried, obtain compound 360 (two total recoverys that go on foot are 67% for 0.010g, 0.028mmol) by flash chromatography on silica gel (10: 1 chloroform/methanol) purifying then.

Reaction scheme 91

It is for the purpose of description that following examples are provided, and limits scope of the present invention never in any form.Application Example

Above-claimed cpd can be used to treatment irritated and asthma, sacroiliitis and/or thrombosis." treatment irritated and asthma, sacroiliitis and/or thrombosis " used herein is meant and is used for the treatment of irritated and asthma, sacroiliitis and thrombosis, and be used for development, bronchoconstriction, the inflammation of Ammonium Glycyrrhizate reaction and cause thrombosis and the formation of the clot of relative disease.The The compounds of this invention of available significant quantity or composition are for example treated allergy, asthma, sacroiliitis or thrombosis among the people warm blooded animal.The method of using antianaphylaxis, anti-asthma, arthritis and the anti-thrombosis drug of significant quantity is known in the art, comprises suction, oral or parenteral admin.Used dosage form includes but not limited to stomach and intestine externally used solution, tablet, capsule, slowly-releasing implant and transdermal drug delivery system; Or the inhalation system of the multiple doses suction apparatus of use Diskus or pressurization.Usually, preferably adopt oral or intravenous administration comes treatment of arthritis and thrombosis, and preferably adopt oral or suction/intranasal administration is treated asthma and allergy.Select dosage and administration frequency to set up effective drug level and do not produce adverse influence.Usually, being used to produce the oral or intravenous administration dosage range that antianaphylaxis, anti-asthma, arthritis or antithrombotic form effect is about 0.01-100mg/kg/ days, is generally 0.1-10mg/kg/ days.Equally, be used to produce in the nose of anti-asthma and antiallergic effect or the inhalation dosage range generally is about 0.01-1mg/kg/ days.

Compound of the present invention or composition and other medicines can be carried out Combined Preparation.For example, can use bronchodilator or glucocorticosteroid is used for the treatment of asthma, uses glucocorticosteroid and be used for the treatment of sacroiliitis or use antihistaminic agent and be used for the treatment of allergy.Also can treat in asthma, allergy, sacroiliitis and the thrombosis one or more with nonsteroidal compound and steroide co-administered of the present invention and/or with nonsteroidal compound and steroide combined utilization of the present invention.

The embodiment of the biologic activity of all cpds described in the synthetic embodiment 1-5 part for example, below is provided.

The anti-thrombosis activity of polyhydroxylated steroide

The present invention finds that described polyhydroxylated steroide of above each several part and intermediate thereof can suppress the caused platelet aggregation of platelet activation factor (PAF).PAF is thrombotic local medium, the formation of prevention clot with treat thrombosis and relevant cardiovascular disorder direct relation arranged.The pilot system that is used to assess the inhibition ability of the platelet aggregation that compound causes the external source sexual stimulus is that antithrombotic forms or the symbol of thrombolysis activity.

From rabbit blood, separate thrombocyte and with 2.4 * 10 ⁸The density of cell/ml adds and contains Ca ²⁺Tyrodes damping fluid (pH7.2) in.Before the stimulation, thrombocyte and each compound are incubated 5 minutes down in 37 ℃.In the presence of each compound, with 1nM platelet activation factor (PAF; EC ₇₅) stimulating platelet and to congregation monitoring 5 minutes.Compound dissolves with methyl-sulphoxide (DMSO), and congregation is recently to measure the percentage of 1nM PAF reaction in the presence of the DMSO of suitable concentration.Control reaction when having DMSO is calculated by the caused inhibition degree of each sample thus as 100%.

Table 1 has provided some can suppress the examples of compounds of the caused platelet aggregation of PAF.

Table 1

All cpds influences * for the rabbit platelet accumulative that stimulates with 0.1nM PAF

	Suppress percentage ratio
	Suppress percentage ratio	Sample number	????80μM
????7	????12.3	Sample number	????80μM
????7	????12.3	????8	????11.6
???165	????51.2	????8	????11.6
???165	????51.2	???236	????100
???241	????93.1	???236	????100
???241	????93.1	???246	?????21
???330	????20.9	???246	?????21

* before stimulating, thrombocyte is incubated 5 minutes with each compound of 80 μ M.The reaction that records for when only having the DMSO of suitable concentration by the inhibition per-cent of the caused reaction of PAF.

Compound discharges the influence of hexosaminidase to rat hypertrophy cell system (RBL-2H3)

Assess its antiallergic activity to what antigen caused from the rat hypertrophy cell system (RBL-2H3) of passive sensitization and the influence of mouse hypertrophy cell system (MC/9) release hexosaminidase by measuring various poly-hydroxy steroide of the present invention.The ability that reagent suppresses mast cell granule composition such as histamine and hexosaminidase release is the active symbol of antianaphylaxis and/or anti-asthma.

In the antigen attack process, discharge hexosaminidase, histamine and other medium from mast cell granule.RBL-2H3 and MC/9 cell are grown in substratum and make it to dinitrophenol (DNP) passive sensitization with Anti-Human DNP (IgE).Cell and each compound (25 μ M) are arised from 37 ℃ of insulations 1 hour, use 0.1mg/ml DNP-HSA (antigen) to stimulate then 15 minutes.Take out supernatant liquor and be used for being determined at the amount of the hexosaminidase that discharges in the antigen attack process.The amount of hexosaminidase is measured by the enzyme catalysis metabolism that detected p-nitrophenyl-N-ethanoyl-β-D-glucosaminide under 410nm in 1 hour with colorimetry in the supernatant liquor.Be used in the effect of each compound that the per-cent of the reaction (subtracting background release) that the antigen that produces when only having DMSO causes records, referring to table 2 and 3.Determine to discharge the inhibition degree of hexosaminidase from cell with these numerical value for what antigen caused.

The rat hypertrophy cell system that table 2 all cpds (25 μ M) causes for antigen from passive sensitization

(RBL-2H3) and mouse hypertrophy cell system (MC/9) discharge the * that influences of hexosaminidase

	Suppress percentage ratio (mean value)
	Suppress percentage ratio (mean value)		Compound number	The RBL-2H3 cell	The MC/9 cell
????333	????69	????71	Compound number	The RBL-2H3 cell	The MC/9 cell
????333	????69	????71	????335	????56	????69
????339	????79	????67	????335	????56	????69
????339	????79	????67	????337	????44	????56
????339	????76	????55	????337	????44	????56
????339	????76	????55	????330	????59	????49
????343	????52	????40	????330	????59	????49
????343	????52	????40	????342	????57	????39
????361	????ND	????13	????342	????57	????39
????361	????ND	????13	????331	????ND	????5
????354	????30	????76	????331	????ND	????5
????354	????30	????76	????346	????16	????61

The ND=undetermined

The rat hypertrophy cell system (RBL-2H3) from passive sensitization that table 3 all cpds (25 μ M) causes for antigen discharges the * that influences of hexosaminidase

	Suppress percentage ratio (mean value) RBL-2H3 cell
	Suppress percentage ratio (mean value) RBL-2H3 cell	????7	?????21.7
???165	?????51.8	????7	?????21.7
???165	?????51.8	???236	??????31
???241	??????22	???236	??????31
???241	??????22	???246	?????29.4
???306	?????40.5	???246	?????29.4
???306	?????40.5	???322	?????25.6
???327	?????35.2	???322	?????25.6
???327	?????35.2	???328	?????34.3
???266	????-12.5	???328	?????34.3
???266	????-12.5	???239	?????9.6
???351	?????33.6	???239	?????9.6
???351	?????33.6	???360	?????76.0

* the reacting phase ratio that produces when only having DMSO of numeric representation, the inhibition per-cent that each compound produces.The influence that the ileum smooth muscle that selected compound causes anaphylactogen shrinks

The ability that the sensitized animal ileum smooth muscle that compound inhibition anaphylactogen causes shrinks is the symbol of antiallergic activity.What adopt when measuring type is the guinea pig ileum of sensitization.Guinea pig ileum is used to assess compound and suppresses the ability that the histamine that causes smooth muscle contraction that anaphylactogen causes and medium discharge.Peritoneal injection 100mg Protalbinic acid and at the 0th day intramuscular injection 50mg Protalbinic acid at the 1st day 50mg of intramuscular injection for the second time Protalbinic acid, makes cavy sensitization then thus.Initial immunity was found animal by sensitization after 21 days, showed as with anaphylaxis occurring after the anaphylactogen attack.Preparation ileum sections is suspended on it in 37 ℃ the Tyrode damping fluid and with containing 5%CO ₂O ₂The contraction of muscle is measured in inflation simultaneously on fore-and-aft plane.The resting tension that tissue is suspended on 2g descends and uses the power conversion transmitter that links with polygraph to measure isometric contraction.To organize histamine to stimulate 3 times to guarantee to produce reproducible contraction with 3 μ M.To organize then with each compound (30 μ M) or 0.15% methyl-sulphoxide (DMSO) in contrast and be incubated 20 minutes, will organize then with the Protalbinic acid of 100 μ g/ml and attack.Protalbinic acid caused shrinkage value in the presence of all cpds is expressed as the percentage ratio of the caused contraction of 3 μ M histamine.Table 4 has been summarized the provide protection that sensitized guinea pig ileum that all cpds causes for Protalbinic acid shrinks.Table 4

The influence that the sensitized guinea pig ileum that all cpds (30 μ M) causes for antigen shrinks.

The contraction that antigen causes is represented * with the percentage ratio of the caused contraction of 3 μ M histamine

Compound number	Suppress percentage ratio
Compound number	Suppress percentage ratio	??????330	??????70.0
??????221	??????60.7	??????330	??????70.0
??????221	??????60.7	??????338	??????64.0
???????7	??????54.0	??????338	??????64.0
???????7	??????54.0	??????333	??????63.0
??????343	??????48.9	??????333	??????63.0
??????343	??????48.9	??????336	??????79.3
??????342	??????28.3	??????336	??????79.3
??????342	??????28.3	??????339	??????40.6
??????335	??????30.7	??????339	??????40.6
??????335	??????30.7	??????337	??????50.7
??????165	??????27.0	??????337	??????50.7
??????165	??????27.0	??????251	????-10.5(stim)
??????361	??????55.0	??????251	????-10.5(stim)
??????361	??????55.0	??????331	??????14.5
??????339	??????36.3	??????331	??????14.5
??????339	??????36.3	??????346	??????62.5
??????334	??????74.0	??????346	??????62.5
??????334	??????74.0	??????266	??????17.4
??????351	??????43.6	??????266	??????17.4
??????351	??????43.6	??????360	??????50.5

* the reacting phase ratio that produced when only having DMSO of numeric representation, the average inhibition percentage ratio that each compound produces, n=3-4.

The influence of the bronchoconstriction that selected compound causes anaphylactogen in vitro and in vivo

The influence of having assessed the bronchoconstriction that multiple compound as herein described causes anaphylactogen is to assess its anti-asthma activity.Compound suppresses in the cavy of sensitization because the ability that the pulmonary function of replying the antigen induction that is produced that antigen is attacked descends is the active symbol of anti-asthma.Specifically, this model can be used for assessing the potential effect in the early stage asthma reaction (EAR) of compound when serious bronchoconstriction appears in treatment.

Cavy is contacted 15 minutes with 1% Protalbinic acid (OA) salt brine solution of atomizing.Find animal after 10 days by sensitization, promptly tracheal tissue attacks further antigen (OA) and produces supersensitivity bronchospasm.Discovery produces reaction from the tracheae of these animals in the mode similar to situation in the body.The preparation tracheal ring also places 37 ℃ Krebs-Henseleit solution, with containing 5%CO ₂O ₂Inflation.The resting tension that tissue is suspended on 2g descends and uses the power conversion transmitter that links with polygraph to measure isometric contraction.To organize with each compound or 0.1% methyl-sulphoxide (contrast) and be incubated 20 minutes, in tissue, add the OA (0.001-100 μ g/ml) of progressive concentration then.After adding last OA concentration and having write down reaction, the mecholyl (can produce maximum tracheae shrink) of tissue with 100 μ M stimulated.OA caused shrinkage value in the presence of all cpds is expressed as the percentage ratio of using the caused maximum collapse of mecholyl (100 μ M).Table 5-7 has summarized the provide protection that tracheal tissue that all cpds causes for OA shrinks.

The isolated tracheal that the selected compound (20 μ M) of table 5 causes anaphylactogen shrinks influences * (research 1)

	???????????????????????????????????????????μg/ml?OA
	???????????????????????????????????????????μg/ml?OA										??0.00 ???1	??0.00 ????3	??0.01	??0.03	??0.1	??0.3	??1.0	?3.0	10.0	??30.0
	?Ctrl	??2.9	??7.7	??11.4	??17.4	??19.7	??26.4	??30.9	??37.2	??43.6	??0.00 ???1	??0.00 ????3	??0.01	??0.03	??0.1	??0.3	??1.0	?3.0	10.0	??30.0	??46.8
?241	?Ctrl	??2.9	??7.7	??11.4	??17.4	??19.7	??26.4	??30.9	??37.2	??43.6	??8.5	??12.6	??17.1	??25.8	??31.0	??33.4	??36.3	??34.4	??35.0	??37.0	??46.8
?241	?236	??1.9	??5.6	??5.6	??5.6	??11.1	??16.7	???39	???28	???39	??8.5	??12.6	??17.1	??25.8	??31.0	??33.4	??36.3	??34.4	??35.0	??37.0	???41
?145	?236	??1.9	??5.6	??5.6	??5.6	??11.1	??16.7	???39	???28	???39	??0.8	??6.2	??5.9	??7.6	??8.3	??13.6	??14.8	??20.0	??28.0	??31.0	???41
?145	?246	??2.6	??4.0	??6.5	??9.7	??12.2	??15.3	??20.0	??21.0	??23.0	??0.8	??6.2	??5.9	??7.6	??8.3	??13.6	??14.8	??20.0	??28.0	??31.0	??22.0

* numeric representation is compared with the caused contraction of mecholyl (100%) of using 100 μ M, the percentage ratio of contraction, and Ctrl=contrasts (0.1%DMSO).

The isolated tracheal that the selected compound (20 μ M) of table 6 causes anaphylactogen shrinks influences * (research 2)

	??????????????????????OAμg/ml
	??????????????????????OAμg/ml					Sample	???0.01	????0.1	????1	????10	????100
????Ctr1	???0.95	????9.0	???25.7	???44.7	???54.6	Sample	???0.01	????0.1	????1	????10	????100
????Ctr1	???0.95	????9.0	???25.7	???44.7	???54.6	????326	????0	???10.25	???23.4	???43.9	???49.1
????327	????0	????0	????9.1	???27.3	???40.9	????326	????0	???10.25	???23.4	???43.9	???49.1

Table 7

The isolated tracheal that compound 330 (30 μ M) causes anaphylactogen shrinks influences * (research 3)

	????????????????????OAμg/ml
	????????????????????OAμg/ml						Sample	???0.001	???0.01	???0.1	????1	????10	????100
???Ctrl	????6.0	???12.0	???26.0	???41.0	???54.0	???59.0	Sample	???0.001	???0.01	???0.1	????1	????10	????100
???Ctrl	????6.0	???12.0	???26.0	???41.0	???54.0	???59.0	????330	????0	???1.90	???9.00	???17.5	???26.0	???30.0

In addition, also measured The compounds of this invention as follows in vivo to influence of Pulmonary Function:

Female Cam Hartley cavy (350-400g) is contacted 15 minutes with 1% Protalbinic acid salt brine solution of atomizing make its sensitization.Find after 10-12 days that animal produces acute allergy to anaphylactogen (Protalbinic acid).Under slight halothane anesthesia, with the 300 μ l PEG solution-treated of animal through port lumen feeding with 300 μ l polyoxyethylene glycol-200 (PEG) or 5mg/kg test compound.Once a day, handled 4 days, 2 hours beginning antigen is attacked after the last administration.Perhaps, compound is carried out inhalation with the Hudson atomisation unit with 6 pounds/square inch oxygen, at the preceding single dose that 50 μ g/kg were provided in 20 minutes of attack.

In surgical procedure, animal is used ketamine (50mg/ml; Intraperitoneal) and xylazine (10mg/kg; Intraperitoneal) and 1% halothane anesthesia.Before being fixed on animal on the body plethysmograph, carrying out tracheotomy and insert water supply oesophagus sleeve pipe.Trachea cannula is connected on the fixed air pipe-in-pipe in the plethysmograph.Use the electrocardiogram monitoring heart function.Animal is used pancuronium bromide (0.8mg/kg; Intramuscular) paralysis and take a breath with the 3ml cheyne stokes respiration with Harvard animalcule air interchanger, frequency is 60 breathings of per minute.Obtain lung resistance and dynamic lung compliance data with the multiple spot analysis from volume, flow and transpulmonary pressure force signal.

In whole experiment, pulmonary function is carried out the resistance and the conformability of continuous monitoring and the mensuration of each time point (for example 0,1,2,3,4,5,10,20 and 30 minute) after antigen is attacked lung.With DIREC physiology logging software data gathering also being used in the physiology mensuration system that computer connects is that the ANADAT software that mechanics of lung mensuration designs is analyzed.This software is from RHT-Info Dat Inc., Montreal, and Qubec, Canada obtains.

After having recorded baseline resistance and conformability, 6 salt solution of animal breath are attacked to it.(variation of pulmonary function should not take place) after 10 minutes, the Protalbinic acid salt brine solution (as the antigenic stimulation thing) of animal breath 6 times 2% or 3% is attacked during this period.The each breathing all carried salt solution and antigen with the Hudson spraying gun.Summarized the provide protection that tracheal tissue that oral administration compound 330 causes for OA shrinks in the following table 8 and 9.

Table 8

Compound 330 (5mg/kg/ days, oral administration 4 days) is in the cavy of sensitization

The influence that the lung resistance that anaphylactogen is caused increases

The timed interval after the attack	Lung resistance (cmH ₂O/ml/sec)
The timed interval after the attack	Lung resistance (cmH ₂O/ml/sec)			Contrast	??????????330
Baseline	????0.287±0.020	????0.275±0.036		Contrast	??????????330
Baseline	????0.287±0.020	????0.275±0.036	?????OA/10s	????0.295±0.024	????0.260±0.036
??????1min	????0.982±0.209	????0.560±0.101	?????OA/10s	????0.295±0.024	????0.260±0.036
??????1min	????0.982±0.209	????0.560±0.101	??????2min	????2.390±0.728	????0.845±0.201
??????3min	????2.627±0.714	????0.887±0.160 ?????(P＜0.06)	??????2min	????2.390±0.728	????0.845±0.201
??????3min	????2.627±0.714	????0.887±0.160 ?????(P＜0.06)	??????4min	????2.801±1.042	????0.778±0.119 ^*
??????5min	????2.514±0.952	????0.791±0.139 ^*	??????4min	????2.801±1.042	????0.778±0.119 ^*
??????5min	????2.514±0.952	????0.791±0.139 ^*	?????10min	????1.329±.209	????0.661±0.141 ^*
?????20min	????1.352±0.494	????0.366±0.046 ^*	?????10min	????1.329±.209	????0.661±0.141 ^*
?????20min	????1.352±0.494	????0.366±0.046 ^*	?????30min	????1.00±0.434	????0.340±0.037 ^*

^*With contrast significant difference is arranged, P＜0.05.

Table 9

The influence that the lung compliance that anaphylactogen is caused reduces

The timed interval after the attack	Lung compliance (ml/cmH ₂O)
The timed interval after the attack	Lung compliance (ml/cmH ₂O)			Contrast	??????????330
Baseline	????0.412±0.053	?????0.31g±0.042		Contrast	??????????330
Baseline	????0.412±0.053	?????0.31g±0.042	?????OA～10s	????0.573±0.083	?????0.435±0.041
????1rnin	????0.077±0.016	?????0.182±0.093	?????OA～10s	????0.573±0.083	?????0.435±0.041
????1rnin	????0.077±0.016	?????0.182±0.093	????2min	????0.029±0.006	?????0.145±0.092
????3min	????0.024±0.003	?????0.133±0.095	????2min	????0.029±0.006	?????0.145±0.092
????3min	????0.024±0.003	?????0.133±0.095	????4min	????0.023±0.003	?????0.124±0.088 ^*
????5min	????0.026±0.002	?????0.125±0.087 ^*	????4min	????0.023±0.003	?????0.124±0.088 ^*
????5min	????0.026±0.002	?????0.125±0.087 ^*	???10min	????0.042±0.002	?????0.1?50±0.082 ^*
???20min	????0.059±0.007	?????0.184±0.061 ^*	???10min	????0.042±0.002	?????0.1?50±0.082 ^*
???20min	????0.059±0.007	?????0.184±0.061 ^*	???30min	????0.077±0.010	?????0.196±0?061 ^*

^*With contrast significant difference is arranged, P＜0.05.

Summarized the provide protection that tracheal tissue that compound 330 inhalations cause for OA shrinks among the following table 10-11.

Table 10 compound 330 (50 μ g/kg suck) is in the cavy of sensitization

The influence that the lung resistance that anaphylactogen is caused increases

After the attack to interbody spacer	Lung resistance (cmH ₂O/ml/sec)
After the attack to interbody spacer	Lung resistance (cmH ₂O/ml/sec)			Contrast	???????????330
Baseline	????0.257±0.019	????????0.300±0.025		Contrast	???????????330
Baseline	????0.257±0.019	????????0.300±0.025	????OA/10s	????0.257±0.046	????????0.288±0.036
????1min	????0.557±0.118	????????0.382±0.033	????OA/10s	????0.257±0.046	????????0.288±0.036
????1min	????0.557±0.118	????????0.382±0.033	????2min	????1.323±0.344	????????0.420±0.044 ^*
????3min	????1.987±0.572	????????0.420±0.051 ^*	????2min	????1.323±0.344	????????0.420±0.044 ^*
????3min	????1.987±0.572	????????0.420±0.051 ^*	????4min	????1.625±0.248	????????0.455±0.047 ^*
????5min	????1.395±0.193	????????0.446±0.124 ^*	????4min	????1.625±0.248	????????0.455±0.047 ^*
????5min	????1.395±0.193	????????0.446±0.124 ^*	???10min	????0.949±0.165	????????0.436=0.036 ^*
???20min	????0.589±0.091	????????0.413±0.076	???10min	????0.949±0.165	????????0.436=0.036 ^*
???20min	????0.589±0.091	????????0.413±0.076	???30min	????0.493±0.067	????????0.412±0.072

^*With contrast significant difference is arranged, P＜0.05.

Table 11

Compound 330 (50 μ g/kg suck) is in the cavy of sensitization

The influence that the lung compliance that anaphylactogen is caused reduces

The timed interval after the attack	Lung compliance (ml/cm H ₂O)
The timed interval after the attack	Lung compliance (ml/cm H ₂O)			Contrast	???????????330
Baseline	????0.515±0.169	??????0.463±0.129		Contrast	???????????330
Baseline	????0.515±0.169	??????0.463±0.129	????OA/1Os	????0.526±0.042	??????0.565±0.062
????1?min	????0.095±0.015	??????0.349±0.059 ^*	????OA/1Os	????0.526±0.042	??????0.565±0.062
????1?min	????0.095±0.015	??????0.349±0.059 ^*	????2?min	????0.044±0.010	??????0.213±0.046 ^*
????3?min	????0.031±0.007	??????0.176±0.045 ^*	????2?min	????0.044±0.010	??????0.213±0.046 ^*
????3?min	????0.031±0.007	??????0.176±0.045 ^*	????4?min	????0.037±0.009	??????0.145±0.046 ^*
????5?min	????0.047±0.007	??????0.146±0.031 ^*	????4?min	????0.037±0.009	??????0.145±0.046 ^*
????5?min	????0.047±0.007	??????0.146±0.031 ^*	????10?min	????0.127±0.053	??????0.138±0.022
????20?min	????0.096±0.009	??????0.193±0.054	????10?min	????0.127±0.053	??????0.138±0.022
????20?min	????0.096±0.009	??????0.193±0.054	????30?min	????0.110±0.010	??????0.181±0.046

^*With contrast significant difference is arranged, P＜0.05.

Summarized the provide protection that tracheal tissue that compound 339 oral administrations cause for OA shrinks among the following table 12-13.

Table 12

Compound 339 (5mg/kg/ days, oral administration 4 days) is in the cavy of sensitization

The influence that the lung resistance that anaphylactogen is caused increases

The timed interval after the attack	Lung resistance (cmH ₂O/ml/sec)
The timed interval after the attack	Lung resistance (cmH ₂O/ml/sec)			Contrast	?????????339
Baseline	????0.25±0.008	????0.249±0.017		Contrast	?????????339
Baseline	????0.25±0.008	????0.249±0.017	????OA/10s	????0.261±0.011	????0.239±0.013
????1?min	????1.781±0.737	????0.326±0.041 ^*	????OA/10s	????0.261±0.011	????0.239±0.013
????1?min	????1.781±0.737	????0.326±0.041 ^*	????2?min	????3.079±1.066	????0.522±0.187 ^*
????3?min	????3.623±0.806	????1.102±0.047 ^*	????2?min	????3.079±1.066	????0.522±0.187 ^*
????3?min	????3.623±0.806	????1.102±0.047 ^*	????4?min	????1.699±0.342	????0.996±0.380
????5?min	????2.783±1.010	????1.014±0.413	????4?min	????1.699±0.342	????0.996±0.380
????5?min	????2.783±1.010	????1.014±0.413	????10min	????1.115±0.348	????0.440±0.099
????20min	????0.624±0.178	????0.296±0.031	????10min	????1.115±0.348	????0.440±0.099
????20min	????0.624±0.178	????0.296±0.031	????30min	????0.465±0.126	????0.291±0.037

^*With contrast significant difference is arranged, P＜0.05.

Table 13

Compound 339 (5mg/1g/ days, oral administration 4 days) is in the cavy of sensitization

The influence that the lung compliance that anaphylactogen is caused reduces

The timed interval after the attack	Lung compliance (ml/cmH ₂O)
The timed interval after the attack	Lung compliance (ml/cmH ₂O)			Contrast	???????.339
Baseline	????0.548±0.116	??????0.463±0.026		Contrast	???????.339
Baseline	????0.548±0.116	??????0.463±0.026	????OA/10s	????0.598±0.129	??????0.442±0.025
????1?min	????0.026±0.005	??????0.172±0.027 ^*	????OA/10s	????0.598±0.129	??????0.442±0.025
????1?min	????0.026±0.005	??????0.172±0.027 ^*	????2?min	????0.018±0.002	??????0.088±0.018 ^*
????3?min	????0.016±0.002	??????0.060±0.017 ^*	????2?min	????0.018±0.002	??????0.088±0.018 ^*
????3?min	????0.016±0.002	??????0.060±0.017 ^*	????4?min	????0.019±0.002	??????0.050±0.013
????5?min	????0.021±0.003	??????0.051±0.011	????4?min	????0.019±0.002	??????0.050±0.013
????5?min	????0.021±0.003	??????0.051±0.011	????10min	????0.043±0.005	??????0.084±0.012 ^*
????20min	????.074±0.007	??????0.123±0.015 ^*	????10min	????0.043±0.005	??????0.084±0.012 ^*
????20min	????.074±0.007	??????0.123±0.015 ^*	????30min	????0.093±0.010	??????0.150±0.012 ^*

^*With contrast significant difference is arranged, P＜0.05.

Summarized the provide protection that tracheal tissue that compound 342 oral administrations cause for OA shrinks among the following table 14-15.

Table 14

Compound 342 (5mg/kg/ days, oral administration 4 days) is in the cavy of sensitization

The influence that the lung resistance that anaphylactogen is caused increases

The timed interval after the attack	Lung resistance (cmH ₂O/ml/sec)
The timed interval after the attack	Lung resistance (cmH ₂O/ml/sec)			Contrast	?????????342
Baseline	????0.214±0.010	??????0.212±0.020		Contrast	?????????342
Baseline	????0.214±0.010	??????0.212±0.020	????OA/10s	????0.204±0.010	??????0.223±0.020
????1?mirn	????2.380±0.83	??????0.453±0.120 ^*	????OA/10s	????0.204±0.010	??????0.223±0.020
????1?mirn	????2.380±0.83	??????0.453±0.120 ^*	????2?min	????4.241±1.04	??????1.786±0.82 ^*
????3?min	????4.657±1.21	??????1.930±0.55 ^*	????2?min	????4.241±1.04	??????1.786±0.82 ^*
????3?min	????4.657±1.21	??????1.930±0.55 ^*	????4?min	????4.088±1.42	??????1.621±0.36 ^*
????5?min	????4.519±1.65	??????1.4816±0.32 ^*	????4?min	????4.088±1.42	??????1.621±0.36 ^*
????5?min	????4.519±1.65	??????1.4816±0.32 ^*	????10min	????1.821±0.38	??????1.002±0.14 ^*
????20min	????0.979±0.23	??????0.524±0.08 ^*	????10min	????1.821±0.38	??????1.002±0.14 ^*
????20min	????0.979±0.23	??????0.524±0.08 ^*	????30min	????0.703±0.24	??????0.354±0.04

^*With contrast significant difference is arranged, P＜0.05.

Table 15

The influence that the lung compliance that anaphylactogen is caused reduces

The timed interval after the attack	Lung compliance (ml/cmH ₂O)
The timed interval after the attack	Lung compliance (ml/cmH ₂O)			Contrast	?????????342
Baseline	????0.441±0.034	??????0.444±0.037		Contrast	?????????342
Baseline	????0.441±0.034	??????0.444±0.037	????OA/10s	????0.509±0.057	??????0.464±0.031
????1?min	????0.028±.007	??????0.154±0.055 ^*	????OA/10s	????0.509±0.057	??????0.464±0.031
????1?min	????0.028±.007	??????0.154±0.055 ^*	????2?min	????0.027±0.012	??????0.073±0.038 ^*
????3?min	????0.016±0.004	??????0.044±0.022 ^*	????2?min	????0.027±0.012	??????0.073±0.038 ^*
????3?min	????0.016±0.004	??????0.044±0.022 ^*	????4?min	????0.017±0.004	??????0.044±0.022
????5?min	????0.018±0.004	??????0.038±0.015	????4?min	????0.017±0.004	??????0.044±0.022
????5?min	????0.018±0.004	??????0.038±0.015	????10min	????0.034±0.004	??????0.048±0.005
????20min	????0.054±0.005	??????0.084±0.005	????10min	????0.034±0.004	??????0.048±0.005
????20min	????0.054±0.005	??????0.084±0.005	????30min	????0.074±0.008	??????0.109±0.006

^*With contrast significant difference is arranged, P＜0.05.

The pulmonary inflammatory influence that selected compound causes anaphylactogen

Inflammatory cell that compound inhibition anaphylactogen causes such as eosinophil and neutrophil's accumulative ability in from the irrigating solution of sensitized animal are the active symbols of anti-asthma.Specifically, this model system can be used for assessing the compounds for treating asthma late phase response effect of (second phase that pneumonia and bronchoconstriction occur).

1ml sterile saline solution by peritoneal injection 1mg Protalbinic acid and 100mg aluminium hydroxide with male Brown Norway rat (200-250g) to egg albumen sensitization.Find after 21 days that animal is to the Protalbinic acid sensitization.Through port lumen feeding is handled medicine for animal or carrier (0.3mlPEF-200), once a day, and totally 4 days.By contacting 60 minutes animal is attacked with the 0.5% Protalbinic acid salt brine solution (producing) of atomizing with the Devillbis spraying gun.Attack and carried out last administration in back 24 hours.Attack after 48 hours, with animal with excessive halothane anesthesia and with lung with 7 * 2ml Sterile Saline lavation (room temperature).The irrigating solution that reclaims placed on ice and centrifugal so that from the supernatant liquor isolated cell with 1200rpm.With cell and of short duration contact of Tris/ ammonium chloride (pH 7.3), in phosphate-buffered saline, wash then to remove red corpuscle.Prepare the centrifugal goods (cytospin) of each cell sample and dye to determine whether to exist the cell that contains peroxidase and to determine eosinophil and neutrophil's quantity.The quantity of inflammatory cell is represented with the percentage ratio of the total cellular score that reclaims in the irrigating solution.Summarized the pulmonary inflammatory provide protection that 330 pairs of anaphylactogens of compound cause in the table 16.

Table 16 compound 330 (5mg/kg/ days, oral administration 4 days) is coming the inflammatory cell that Protalbinic acid causes

Accumulative influences # in the Brown of sensitization Norway rat irrigating solution

	The percentage ratio of the total cell that in irrigating solution, reclaims
				Handle	The cell that peroxidase stain is positive	Eosinophil	The neutrophil(e) cell
Contrast	???0.55±0.27	?0.69±0.30	???0.665±0.31	Handle	The cell that peroxidase stain is positive	Eosinophil	The neutrophil(e) cell

Only use OA	??36.03±5.55	???20.00±2.65	??11.58±1.53
Only use OA	??36.03±5.55	???20.00±2.65	??11.58±1.53	Compound 330+OA	??5.65±2.44 ^*	???1.98±0.78 ^*	??6.16±4.54

# carries out administration with 300 μ l polyoxyethylene glycol-200 as carrier with medicine.The animal of treated with medicaments is not only accepted 300 μ l polyoxyethylene glycol.

^*With contrast significant difference is arranged, P＜0.05.

The selected effect of compound in supersensitivity sheep asthmatic model

Studied the effect of selected compound in supersensitivity sheep asthmatic model.

Adopting the supersensitivity sheep model is because this model shows the principal feature relevant with asthma.This model shows natural allergy, early stage (acute) bronchoconstriction, late period bronchoconstriction, pneumonia and bronchial overreaction.Animal regains consciousness in this model, the animal autonomous respiration, thus can measure the bronchoconstriction and the acute airway overreaction of airway.

Endotracheal tube and foley's tube are inserted into esophagus hypomere to the sheep (30-40kg) of the natural sensitization of Ascaris suum.Measure pleura pressure with esophageal intubation, and with deeply and be positioned the side-holes catheter mensuration pressure afterwards of endotracheal tube far-end.Poor with the transpulmonary pressure power between differential pressure sensor tube systems measurement tracheae and pleura pressure.

The near-end of endotracheal tube is connected with the Fleisch pneumotachograph to measure the change of flow.From the pressure measuring value of transpulmonary pressure power, respiratory volume (obtaining) and flow from the numerical integration of flow signal by bit traffic (mid-flow) technique computes go out lung resistance (R _L).SR _LCalculating formula be R _L/ V _Tg(V _Tg=thorax gas volume).

Produce in aerosol and importing and Havard respiratory organ and the placed in-line T shape of the trachea cannula parts with disposable spraying gun.With the dosage of dosimetry system control aerosol, described dosimetry system is made up of magnetic valve and pressurized air (20 pounds/square inch), and pressurized air discharges when the beginning of each imbibition cycle.With aerosol with the frequency administration of the damp formula tolerance of 500ml with 20Hz.

The compound that to select from the present invention is dissolved in DMSO with the form of storing solution and dilutes with salt solution.At preceding 30 minutes of attack and the back compound of using 400 μ g/kg in 4 hours to animal of attack; Perhaps, use the compound of 4 days 400 μ g/kg to animal, last administration was carried out in attack in preceding 2 hours.To be diluted to concentration in phosphate-buffered saline be 82000 PNUs/ml and passed through aerosol drug delivery in 20 minutes with Ascaris suum extract.Carbachol is dissolved in PBS, forms the concentration of 0.25,0.5,1.0,2.0 and 4.0% (weight/volume).In whole research process, each animal is all as self contrast.

Antigen is attacked the back and was measured once than lung resistance (SR in per 60 minutes _L) and continue 8 hours.The initial attack measured the overreaction of airway to carbachol after 24 hours.

Table 17-18 has summarized compound 330 acute administrations and (had attacked preceding 30 minutes and attacked back 4 hours; 400 μ g/kg) contrast lung resistance and over-reactive provide protection.

Table 17 compound 330 (was attacked preceding 30 minutes and was attacked back 4 hours; 400 μ g/kg, inhalation) the ratio lung resistance that in the sheep of Ascaris suum sensitization anaphylactogen is caused changes influences #

The timed interval after the attack	Than lung resistance (% baseline)
The timed interval after the attack	Than lung resistance (% baseline)			Contrast	????330
Baseline	????7±10	????2±5		Contrast	????330
Baseline	????7±10	????2±5	????-0.5	????7±10	????0±4
Attack (0)	???266±33	????268±35	????-0.5	????7±10	????0±4
Attack (0)	???266±33	????268±35	????1	???197±51	????117±11
????2	???77±21	????49±11	????1	???197±51	????117±11
????2	???77±21	????49±11	????3	???68±36	????32±6
????4	???23±7	????20±6	????3	???68±36	????32±6
????4	???23±7	????20±6	????5	???73±12	????14±4 ^*
????6	??132±29	????14±5 ^*	????5	???73±12	????14±4 ^*
????6	??132±29	????14±5 ^*	????6.5	??126±15	????18±6 ^*
????7	??129±16	????10±2 ^*	????6.5	??126±15	????18±6 ^*
????7	??129±16	????10±2 ^*	????7.5	??156±13	????17±8 ^*
????8	??123±27	?????5±3 ^*	????7.5	??156±13	????17±8 ^*

# drug administration amount is 3ml (salt brine solution of 66%DMSO).Carrier itself does not have influence.Animal was respectively handled once in preceding 30 minutes of attack and attack in back 4 hours. ^*With contrast significant difference is arranged, P＜0.05.

Table 18 compound 330 (was attacked preceding 30 minutes and was attacked back 4 hours; 400 μ g/kg, inhalation) in the sheep of Ascaris suum sensitization for segmental bronchus to the over-reactive # that influences of carbachol

	????PC ₄₀₀(breathing unit) hyperreactive
	????PC ₄₀₀(breathing unit) hyperreactive			Contrast	???????330
Baseline	?26.65±3.08	??26.01±3.06		Contrast	???????330
Baseline	?26.65±3.08	??26.01±3.06	After the attack	?12.28±1.49	??22.79±6.11 ^*

# drug administration amount is 3ml (salt brine solution of 66%DMSO).Carrier itself does not have influence.Animal was respectively handled once in preceding 30 minutes of attack and attack in back 4 hours.The initial overreaction of measuring after 24 hours carbachol of attacking.

^*With contrast significant difference is arranged, P＜0.05.Table 19-20 has summarized and has used (400 μ g/kg) compound 330 contrast lung resistance and over-reactive provide protections in 4 days.Table 19 compound 330 (400 μ g/kg, inhalation 4 days) is right in the sheep of Ascaris suum sensitization

Ratio lung resistance that anaphylactogen causes changes influences #

The timed interval after the attack	Than lung resistance (% baseline)
The timed interval after the attack	Than lung resistance (% baseline)			Contrast	???????????330
Baseline	?????2.00±2.00	?????5.25±4.50		Contrast	???????????330
Baseline	?????2.00±2.00	?????5.25±4.50	????-0.5	?????2.00±2.00	?????-3.75±4.29
Attack (0)	????248.25±85.71	????126.00±19.11	????-0.5	?????2.00±2.00	?????-3.75±4.29
Attack (0)	????248.25±85.71	????126.00±19.11	????1	????170.75±58.62	????34.00±10.75
????2	????74.25±17.10	????11.50±6.18 ^*	????1	????170.75±58.62	????34.00±10.75
????2	????74.25±17.10	????11.50±6.18 ^*	????3	????82.00±6.82	????-15.00±37.67 ^*
????4	????21.25±5.41	??????4.00±1.78 ^*	????3	????82.00±6.82	????-15.00±37.67 ^*
????4	????21.25±5.41	??????4.00±1.78 ^*	????5	????57.50±7.51	??????-4.50±4.17 ^*
????6	???132.00±9.68	??????7.75±9.75 ^*	????5	????57.50±7.51	??????-4.50±4.17 ^*
????6	???132.00±9.68	??????7.75±9.75 ^*	????6.5	???153.75±21.93	???????5.50±4.87 ^*
????7	????173.75±21.74	??????10.75±4.91 ^*	????6.5	???153.75±21.93	???????5.50±4.87 ^*
????7	????173.75±21.74	??????10.75±4.91 ^*	????7.5	????148.00±20.96	??????4.00±2.35 ^*
????8	????124.75±28.53	???????3.25=4.73 ^*	????7.5	????148.00±20.96	??????4.00±2.35 ^*

# drug administration amount is 3ml (salt brine solution of 66%DMSO).Carrier itself does not have influence.Animal was handled 4 days, and last administration was carried out in attack in preceding 30 minutes.

^*With contrast significant difference is arranged, P＜0.05.

Table 20 compound 330 (400 μ g/kg/ days, inhalation 4 days) is in the sheep of Ascaris suum sensitization

For segmental bronchus to the over-reactive # that influences of carbachol

	????PC ₄₀₀(breathing unit) hyperreactive
	????PC ₄₀₀(breathing unit) hyperreactive			Contrast	330
Baseline	?25.1±1.54	??21.26±2.75		Contrast	330
Baseline	?25.1±1.54	??21.26±2.75	After the attack	?11.9±1.09	???21.21±3.10 ^*

# drug administration amount is 3ml (salt brine solution of 66%DMSO).Carrier itself does not have influence.Animal was handled 4 days, and last administration was carried out in attack in preceding 30 minutes.The initial overreaction of measuring after 24 hours carbachol of attacking.

^*With contrast significant difference is arranged, P＜0.05.

Selected compound is to the influence of the transcription factor that relates to inflammatory process

The characteristics of many chronic inflammatory diseases are activation of the gene of some known maintenance inflammatory conditions.Comprising cytokine, chemokines, adhesion molecule, transcription factor and proteolytic enzyme.What play central role in the expression of molecule before multiple these inflammation is the protein that a class is called transcription factor.Known is NF-κ B for the very important a kind of transcription factor of state before the inflammation.Many chronic diseases raise relevant with activated NF-κ B level.Described disease comprises atherosclerosis, cancer, infectious diseases and various inflammation disease, comprises asthma, inflammatory bowel disease, sacroiliitis, local asphyxia/pour into again and the inflammatory skin disease.Found that compound of the present invention can suppress the activation of the caused NF-κ B of Buddhist ripple ester (NF-κ B activator).

The employing gel shift is analyzed, and detecting selected The compounds of this invention by mensuration NF-κ B and DNA specificity site bonded level influences NF-κ B activated.Be used to measure NF-κ B bonded oligonucleotide and carry out mark by the following method.It is 20 μ l that 5 μ l NF-κ B oligonucleotide (8.9pmol), 2 μ l 10x T4-polynucleotide kinase damping fluids, 10 T4-of unit polynucleotide kinases and 1 μ l γ-P-32-dATP (10 μ Ci) water are added to final volume.This reaction solution is incubated 30 minutes in 37 ℃.Use 2 μ l0.5M EDTA and 2 μ l 3M sodium acetate (pH5.2) termination reactions then.Add 100% ethanol of 2.5 times of volumes and with the mixture that forms centrifugal 10 minutes in 15000g (Eppendorf tube).With settling for several times, dried under the room temperature 10 minutes then, be suspended in again then (ultimate density is 0.75pmol/2 μ l) in the distilled water with 70% washing with alcohol.Cell (RBL-2H3 and A-549) is washed 2 times under room temperature with phosphate-buffered saline (PBS).Scrape 5ml PBS and centrifugal (1500rpm, room temperature, Beckman GPR whizzer) from the tissue culture dish it with the cell spatula.After removing supernatant liquor, cell is suspended in 2 times of buffer A to volume of sediment (0.25M sucrose, 20mM Hepes (pH7.9), 10mM KCl, 1.5mM MgCl again ₂, 0.5mM DTT, 0.5mM spermidine, 0.15mM spermine) in.With its recentrifuge and with cell with 10 ⁸The concentration of cell/ml is suspended in the identical damping fluid again.With insulation under the cell room temperature 5 minutes.It is that 400 μ g/ml (4 μ l/100 μ l buffer A) also are no more than 90 seconds with the insulation under reversing gently of this suspension that adding lysolecithin (solution of 10mg/ml in buffer A) makes ultimate density.The ice-cold buffer A that the adding two volumes contains 3%BSA stops cytolysis rapidly.Under 4 ℃, collected nuclear in centrifugal 1 minute with Eppendorf tube with 4000rpm.Remove supernatant liquor and settling be suspended in the buffer A that contains 3%BSA again, then under 4 ℃ with centrifugal 60 seconds of 30000g (Beckman, TL-100).To examine with about 10 ⁷The concentration of nuclear/ml is suspended in ice-cold buffer B (20mM Hepes (pH 7.9), 25% (volume) glycerine, 0.6M KCl, 1.5mM MgCl again ₂, 0.2mM EDTA, 0.5mM DTT, 0.5mMPMSF) in.To examine destruction (40% intensity setting, MICROSON, ultrasonic cell disintegration machine) in the pulse with 2x5 second on ice by supersound process.Homogenate was stirred gently on ice 30 minutes, centrifugal with Beckman TL-100 down with 25000g then at 4 ℃.Take out supernatant liquor then,, then preserve down in-70 ℃ as not using immediately.The combination of measuring NF-κ B DNA as follows is active: with 2 μ l 10x binding buffer liquid { 20mM HEPES (pH 7.5), 50mM KCl, 5mM MgCl ₂, 200 μ g/mg BSA (Sigma # B-2185), 8% glycerine }, 0.4 μ lPolydI-dC (0.5mg/ml storing solution), 2.0 μ l ³²The oligonucleotide of P-mark mixes from the isolated protein of nucleus with 5 μ g.With distilled water the mixture that forms being transferred to final volume is 20 μ l.Then it is incubated 5 minutes to carry out combination on ice.Continue insulation (20-30 minute) at room temperature.Then with sample on the sample to 4.5% acrylamide gel { 6ml (29: 1) acrylamide: bis, 2ml 5x tbe buffer liquid, 800 μ l, 50% glycerine, 31ml distilled water 150 μ l 10%APS (ammonium persulphate), 40 μ lTEMED}.With acrylamide gel pre-deployment 1.5 hours in 0.25x tbe buffer liquid (10 volts/cm), exchange buffering liquid at last sample and before launching real sample then.

Table 21 has provided the influence to the NF-kB activity of the selected The compounds of this invention that records by the gel shift binding analysis.

Table 21

Selected compound is using TPA (0.1 μ M) to stimulate

In the RBL-2H3 cell NF-κ B bonded is influenced #

Handle	Inhibition per-cent (0.1 μ M) to the TPA reaction
Handle	Inhibition per-cent (0.1 μ M) to the TPA reaction	165(10μM)	???????????????54
330(1μM)	???????????????66	165(10μM)	???????????????54
330(1μM)	???????????????66	333(1μM)	???????????????34
339(1μM)	???????????????65	333(1μM)	???????????????34

Before # stimulates with TPA, with compound or carrier (0.1%DMSO) and cell (RBL-2H3) pre-incubation 2 hours.Cell is stimulated 2.5 hours to activate NF-κ B with 0.1 μ M TPA.All numerical value are all represented with the inhibition percentage ratio of contrast (stimulating with 0.1 μ M TPA in the presence of carrier).

All publications and the patent application mentioned in this specification sheets all are incorporated herein by reference, and it is identical that it quotes the degree that degree and each publication or patent application quote separately as a reference.

Although below for the purpose of description specific embodiments of the present invention is described, clearly can carries out various changes and do not exceed the spirit and scope of the invention.

Claims

1, a kind of compound of following formula:

Comprise its pharmacy acceptable salt and solvate, wherein:

C17 is by (c), (d), (e), (f), (g), (h) with any replacement (i):

(c)=C (R ²) (R ³), but C14 during by methyl substituted except;

(d)-R ⁵With-OR ⁶, as long as A ring and B ring are not aromatic ring, and working as C10 by methyl substituted, C5 directly is not connected with oxygen, wherein R ⁵And R ⁶Can form direct bond together, thereby C17 is carbonyl, perhaps R ⁵And R ⁶Form 3-6 unit's cyclic ethers or 4-6 membered ring lactone with C17; Otherwise, R ⁵Be R ⁴Or-OR ⁶, R ⁶Be R ¹Or R ⁴

ⅰ) C5 is replaced by hydrogen with the α configuration, and C3 is not connected with oxygen; And replace with two hydrogen atoms as C3, then C17 is not by-CH (CH ₃) (CH ₂) ₃CH (CH ₃) ₂Or-CH (CH ₃) (CH ₂) 2C (=O) OCH ₃Replace;

ⅱ) C10 and C13 be simultaneously by methyl substituted, and as C10 during by methyl substituted, then C14 is not by methyl substituted, and the A ring is not an aromatic ring;

ⅵ) C3 and C4 all are connected with identical Sauerstoffatom, thereby form an oxyethane ring, and condition is: when C5 have hydroxyl or-OR ¹During substituting group, C7 does not have the carbonyl substituted base;

(g) ring structure of following formula

(h) two hydrogen atoms are not as long as C3 is by carbonyl substituted;

X represents fluorine, chlorine, bromine and iodine.

2, according to the compound of claim 1, this compound has following formula

Comprise its pharmacy acceptable salt and solvate, wherein:

C14 is-X ,-OR ¹Or except that methyl-R ⁴

X represents fluorine, chlorine, bromine and iodine.

3, according to the compound of claim 1, this compound has following formula

Comprise its pharmacy acceptable salt and solvate, wherein:

A, B, C and D ring can be complete saturated rings, fractional saturation ring or complete unsaturated ring independently, and condition is that A ring and B ring all are not aromatic ring;

X represents fluorine, chlorine, bromine and iodine.

4, according to the compound of claim 1, this compound has following formula

Comprise its pharmacy acceptable salt and solvate, wherein:

C3 quilt=C (R ⁴) (R ⁴) and-C (R ⁴) (R ⁴) (C (R ⁴) (R ⁴)) _n-one of replace, wherein n is 1 to about 6, perhaps by-X and-R ⁴In two replacements, condition is that C3 is not connected with Sauerstoffatom, and when C3 is replaced by two hydrogen atoms, then C17 is not by-CH (CH ₃) (CH ₂) ₃CH (CH ₃) ₂Or-CH (CH ₃) (CH ₂) ₂C (=O) OCH ₃Replace;

X represents fluorine, chlorine, bromine and iodine.

5, according to the compound of claim 1, this compound has following formula

Comprise its pharmacy acceptable salt and solvate, wherein:

X represents fluorine, chlorine, bromine and iodine.

6, according to the compound of claim 1, this compound has following formula

Comprise its pharmacy acceptable salt and solvate, wherein:

Condition is that C17 is not by any replacement in the following radicals:

C8 quilt-X or-R ⁴Replace, preferably directly be not connected with oxygen;

X represents fluorine, chlorine, bromine and iodine.

7, according to the compound of claim 1, this compound has following formula

Comprise its pharmacy acceptable salt and solvate, wherein:

G is-C (=O)-,-CH (OR ¹)-,-C (R ⁴) (OR ¹)-or-C (OR ¹) (OR ¹)-;

X represents fluorine, chlorine, bromine and iodine.

8, according to the compound of claim 1, this compound has following formula

Compound also comprises its pharmacy acceptable salt and solvate, wherein:

X represents fluorine, chlorine, bromine and iodine;

9, according to the compound of claim 1, this compound has and is selected from following structural formula:

Comprise its pharmacy acceptable salt and solvate, wherein:

C5 is replaced by hydrogen atom;

C17 quilt (c), (d), (e) or (f) replacement:

(e) at least a substituting group that contains Sauerstoffatom, it is selected from=O ,-(O (C (R ⁴) (R ⁴)) _nO)-,-OH and-OR ¹, wherein, n is 1-about 6;

R ¹Be H, thereby or a kind of blocking group make-OR ¹Be the oh group of protection, wherein, be attached on the adjacent carbons-OR ¹Group can form the ring structure of two hydroxyls of protection together;

Each R ⁴All be independently selected from H and R ⁵

X represents fluorine, chlorine, bromine and iodine.

10, according to each compound of claim 1-9, it has following formula

11, according to each compound of claim 1-9, it has following formula

12, according to each compound of claim 1-9, wherein C10 and C13 are respectively by methyl substituted.

13, according to each compound of claim 1-9, wherein C11 and C12 are only replaced by hydrogen respectively.

14, according to each compound of claim 1-9, wherein C17 is selected from C ₁-C ₇The substituting group of alkyl replaces.

15, according to each compound of claim 1-9, wherein C17 is selected from formula=C (C ₁-C ₅) ₂With=CH (C ₁-C ₆) substituting group replace.

16, according to each compound of claim 1-9, wherein the C17 substituting group of hydroxyl that is selected from carbonyl, hydroxyl and the protection of carbonyl, protection replaces.

17, according to each compound of claim 1-9, wherein the C3 substituting group that is selected from the hydroxyl of halogen, hydroxyl and protection replaces.

18, according to each compound of claim 1-9, wherein C3 quilt-G-R ⁶Replace, wherein, G is selected from direct bond, O, S or NH, R ⁶Be aryl, alkyl, aralkyl and alkaryl, they are replaced by at least one hydrophilic residue, and described hydrophilic residue is selected from sulfate radical, phosphate radical, carboxylate radical, nitro, ammonium, polyoxyalkylene and sugar, and described compound comprises its pharmacy acceptable salt.

19, according to each compound of claim 1-9, wherein C3 and C4 are all replaced by oxygen, and form epoxide together, acetal or ketal.

20, according to each compound of claim 1-9, wherein A, B, C and D ring are saturated rings.

21, according to each compound of claim 1-9, wherein the A ring is unsaturated ring.

22,, wherein, between C4 and C5, there are two keys according to each compound of claim 1-9.

23, according to each compound of claim 1-9, wherein C6 and C7 are all replaced by hydrogen.

24, according to each compound of claim 1-9, wherein C6 and C7 are all replaced by hydroxyl.

25, according to each compound of claim 1-9, wherein R ⁵Be C ₁-C ₃₀Alkyl.

26, a kind of pharmaceutical composition, it comprises each compound and pharmaceutically acceptable carrier or thinner of aforesaid right requirement 1-25.

27, a kind of pharmaceutical composition, it comprises a kind of compound and pharmaceutically acceptable carrier or thinner, and this compound has following formula

Comprise its pharmacy acceptable salt and solvate, wherein:

(b) representative-X ,-R ⁴With-OR ¹In two, each is selected independently;

Each R ⁴All be independently selected from H and R ⁵

X represents fluorine, chlorine, bromine and iodine;

Condition is that C15 does not link to each other with Sauerstoffatom.

28, according to the composition of claim 27, wherein C17 is replaced by alkyl.

29, according to the composition of claim 27, wherein C17 is by C ₁-C ₇Alkyl replaces.

30, according to the composition of claim 27, wherein C17 is by formula=C (R ⁴) (R ⁴) olefin group replace.

31, according to the composition of claim 27, R wherein ⁴Be hydrogen or C ₁-C ₁₀Alkyl.

32, according to the composition of claim 27, wherein alkyl does not comprise

33, according to the composition of claim 27, wherein C17 is replaced by two atoms that are independently selected from hydrogen and halogen atom.

34, according to the composition of claim 27, wherein C17 is replaced by at least one Sauerstoffatom.

35, according to the composition of claim 27, wherein C17 is replaced by the hydroxyl of hydroxyl or protection.

36, according to the composition of claim 27, wherein C17 is by the carbonyl substituted of carbonyl or protection.

37, according to the composition of claim 27, wherein the C17 alkoxy replaces.

38, according to the composition of claim 27, wherein the substituting group on the C17 does not comprise

39, according to the composition of claim 27, wherein C15 is replaced by two hydrogen atoms.

40, according to the composition of claim 27, wherein C4 by hydrogen and-X ,-R ⁵Or-OR ¹One of replace.

41, according to the composition of claim 27, wherein C5 is replaced by hydrogen.

42, according to the composition of claim 27, wherein C4 links to each other with at least one hydrogen, and when C4 linked to each other with two hydrogen atoms, C3 did not link to each other with Sauerstoffatom, does not also link to each other with two hydrogen atoms.

43, according to the composition of claim 27, wherein, only work as C3 and promptly do not link to each other with Sauerstoffatom, when not linking to each other with two hydrogen atoms again, C4 just links to each other with two hydrogen atoms, and only when C4 did not link to each other with two methyl or formyl radical, C4 just linked to each other with methyl.

44, according to the composition of claim 27, wherein the hydrogen at C5 has the α configuration.

45, according to the composition of claim 27, wherein C6-OR ¹Has the α configuration.

46, according to the composition of claim 27, wherein C7-OR ¹Has beta comfiguration.

47, according to the composition of claim 27, wherein C6-OR ¹Have the α configuration, C7-OR ¹Has beta comfiguration.

48, according to the composition of claim 27, wherein at least one among C3 and the C4 links to each other with Sauerstoffatom.

49, according to the composition of claim 27, wherein C3 all links to each other with Sauerstoffatom with C4.

50, according to the composition of claim 27, wherein C10 is by methyl substituted.

51, according to the composition of claim 27, wherein C13 is by methyl substituted.

52, according to the composition of claim 27, wherein C10 and C13 are all by methyl substituted.

53, according to the composition of claim 27, wherein C6 all links to each other with hydrogen atom with C7.

54, according to the composition of claim 27, wherein at least one is only replaced by hydrogen atom among C1, C2, C3, C4, C5, C8, C9, C10, C11, C12, C13, C14, C15, C16 and the C17.

55, according to the composition of claim 27, wherein C1 and C2 are only replaced by hydrogen atom.

56, according to the composition of claim 27, wherein C11 and C12 are only replaced by hydrogen atom.

57, according to the composition of claim 27, wherein C15 and C16 are only replaced by hydrogen atom.

58, according to the composition of claim 27, wherein A, B, C and D ring are saturated rings.

59, according to the composition of claim 27, wherein the A ring does not comprise twin nuclei.

60, according to the composition of claim 27, wherein C3 and C4 are not only replaced by hydrogen atom simultaneously.

61, a kind of method for the treatment of asthma comprises each the compound or its salt of the claim 1-25 that needs the patient of this treatment significant quantity.

62, a kind of method for the treatment of asthma comprises the pharmaceutical composition of the claim 26 that needs the patient of this treatment significant quantity.

63, a kind of method for the treatment of asthma comprises each the pharmaceutical composition of claim 27-60 that needs the patient of this treatment significant quantity.

64, a kind of treatment method hypersensitive comprises each the compound or its salt of the claim 1-25 that needs the patient of this treatment significant quantity.

65, a kind of treatment method hypersensitive comprises the pharmaceutical composition of the claim 26 that needs the patient of this treatment significant quantity.

66, a kind of treatment method hypersensitive comprises each the pharmaceutical composition of claim 27-60 that needs the patient of this treatment significant quantity.

67, a kind of method of treatment of arthritis comprises each the compound or its salt of the claim 1-25 that needs the patient of this treatment significant quantity.

68, a kind of method of treatment of arthritis comprises the pharmaceutical composition of the claim 26 that needs the patient of this treatment significant quantity.

69, a kind of method of treatment of arthritis comprises each the pharmaceutical composition of claim 27-60 that needs the patient of this treatment significant quantity.

70, the thrombotic method of a kind of treatment comprises each the compound or its salt of claim 1-25 that needs the patient of this treatment significant quantity.

71, the thrombotic method of a kind of treatment comprises the pharmaceutical composition of the claim 26 that needs the patient of this treatment significant quantity.

72, the thrombotic method of a kind of treatment comprises each the pharmaceutical composition of claim 27-60 that needs the patient of this treatment significant quantity.

73, a kind of treatment and patient NF _KThe method of the illness that the rising of B activity level is relevant comprises needing the patient of this treatment to reduce NF _KEach compound or its salt of the claim 1-25 of the active significant quantity of B.

74, a kind of treatment and patient NF _KThe method of the illness that the rising of B activity level is relevant comprises needing the patient of this treatment to reduce NF _KThe composition of the claim 26 of the active significant quantity of B.

75, a kind of treatment and patient NF _KThe method of the illness that the rising of B activity level is relevant comprises needing the patient of this treatment to reduce NF _KEach pharmaceutical composition of the claim 27-60 of the active significant quantity of B.

76, a kind of to 6, introduce the method for the outer olefin group of ring in the 7-titanium dioxide steroide, comprise the compound that a kind of formula (10) are provided, the Witting reagent of formula (10) compound and formula (11) is reacted in the presence of alkali, obtain the olefin(e) compound of a kind of formula (12)

Wherein, each compound of formula (10) and formula (12) comprises its pharmacy acceptable salt and its solvate, wherein:

77, according to the method for claim 76, wherein alkali is selected from sodium tert-butoxide, potassium tert.-butoxide and sodium hydride, and alkali mixes with aprotic solvent, and described solvent comprises toluene, tetrahydrofuran (THF), CH ₂Cl ₂, dimethyl formamide, methyl-sulphoxide, benzene and ether.

78, according to the method for claim 76, wherein Ra and Rb are independently selected from hydrogen and C ₁-C ₇Alkyl, X are selected from chlorine, bromine and iodine.

79, a kind of with 6 α, 7 β-titanium dioxide group is introduced the method in the steroide, comprise a kind of steroide that has carbonyl in the C7 position and have the formula (13) of two keys at C5 and C6 interdigit is provided, with carbonyl reduction is hydroxyl, with two key hydroborations, obtain hydroxyl again, thereby the hydroxyl of C6 position has α-configuration in the C6 position, the hydroxyl of C7 position has beta configuration

X represents fluorine, chlorine, bromine and iodine.

80, according to the method for claim 79, wherein reduction reaction adopts sodium borohydride and the combination of Cerium II Chloride (III) hexahydrate to finish.

81, according to the method for claim 79, wherein hydrogenation adopts a kind of hydrogenant agent, and it is selected from BH ₃And 9-BBN, hydrogenation carries out in aprotic solvent, and described solvent comprises tetrahydrofuran (THF), CH ₂Cl ₂, ether, methyl-sulphoxide and dithiocarbonic anhydride, and then handle with superoxide that is selected from hydrogen peroxide and tertbutyl peroxide and the alkali that is selected from sodium hydroxide and potassium hydroxide.

82, a kind of method that is used for introducing hydroxyl in the C3 position of steroid nucleus stereoselectivity, this method comprises provides a kind of compound that has the formula (15) of carbonyl on the C3 position, with reductive agent carbonyl reduction is become hydroxyl, thus at least a in the formula of obtaining (16) and (17) compound

Wherein, each of formula (15), (16) and (17) compound comprises its pharmacy acceptable salt and its solvate, wherein:

X represents fluorine, chlorine, bromine and iodine.

83,2 method according to Claim 8, wherein reductive agent is selected from three amyl group lithium borohydrides, 3-sec-butyl lithium borohydride, three sec-butyl POTASSIUM BOROHYDRIDE, mainly to obtain the oxy-compound of formula (16).

84,2 method according to Claim 8, wherein reductive agent is selected from sodium borohydride and LiAlH ₄, mainly to obtain the oxy-compound of formula (17).

85,2 method according to Claim 8, wherein the ratio of reduction back formula (16) compound and formula (17) compound is not 1: 1.