TW593305B - Processes for preparation of 9,11-epoxy steroids and intermediates useful therein - Google Patents

Abstract

Multiple novel reaction schemes, novel process steps and novel intermediates are provided for the synthesis of epoxymexrenone and other compounds of formula I, wherein: -A-A- represents the group -CHR4-CHR5- or -CR4=CR5-; R3, R4 and R5 are independently selected from the group consisting of hydrogen, halo, hydroxy, lower alkyl, lower alkoxy, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, cyano, aryloxy, R1 represents an alpha-oriented lower alkoxycarbonyl or hydroxyalkyl radical, -B-B- represents the group -CHR6-CHR7- or an alpha- or beta-oriented group, where R6 and R7 are independently selected from the group consisting of hydrogen, halo, lower alkoxy, acyl, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxycarbonyl, acyloxyalkyl, cyano, aryloxy, and R8 and R9 are independently selected from the group consisting of hydrogen, halo, lower alkoxy, acyl, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxycarbonyl, acyloxyalkyl, cyano, aryloxy, or R8 and R9 together comprise a carbocyclic or heterocyclic ring structure, or R8 or R9 together with R6 or R7 comprise a carbocyclic or heterocyclic ring structure fused to the pentacyclic D ring, wherein the intermediates include compounds of formula D, E, F, 211, 210, 209, 208, 207, 206, 205, 204 and 203 as defined in claims 34 to 57.

Description

593305 5. Description of the invention (BACKGROUND OF THE INVENTION The present invention relates to a new class of methods for the preparation of 9, u_epoxy steroids, especially those of the 20 spiroxo (sounding coffee) series and their analogs, used in children steroids Novel intermediates in the preparation of compounds, and methods for preparing the intermediates. Most particularly, the present invention is directed to the preparation of 9- "epoxy] 7α-Aceto-3_oxopregnancy_4_ene ~ , 2-methylene chloride dicarboxylate, r-lactone (eplerenone; epoxy mexenolide): and an advantageous method. Preparation is described in US Patent No. 4,559,332 20 · Spiro-oxygen-based compound. The compound produced according to the method of 332 patent has an oxygen ring that is open in the formula E: Please read the precautions on the back before filling in this Page 』Φ

IA-Ordered by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs -AA- represents the group -CH2-CH2- or _CH = CH-, R1 represents a low-carbon alkoxycarbonyl or hydroxycarbonyl group, -B_Β -Represents a group -CH2-CH2- or a group of orientation, ^ CH-

-CH • C Η * CH-

III -4- This paper size is in accordance with Chinese National Standard (CNS) A4 specification (210X 297 mm) 593305 Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7 ____ B7 V. Description of the invention (2) R6 and R7 are hydrogen, X Represents two hydrogen atoms or oxygen, γ1 and Y2 together represent oxygen _〇_, or Y1 represents a hydroxyl group, and Y2 represents a hydroxyl group, a low-carbon alkoxy group, or if χ represents H2, a low-carbon alkane can also be used. Oxygen, and salts of such compounds, where χ represents oxygen and ¥ 2 represents a light group, which is equivalent to 17 /? Hydroxy_2ΐ-carboxylic acid. Second Earth U.S. Patent No. 4,559,332 describes a number of methods for preparing epoxy Mexley and related compounds of formula 1A. The advent of new epoxy resins and expansion of clinical use (Mexico Reynolds) has created a need for improved methods of manufacturing this and other related steroids. SUMMARY OF THE INVENTION The primary objective of the present invention is to provide the preparation of epoxy resins with common structural characteristics Improved methods of mexanol, other 20-spiroxane, and other related steroids. Among the specific objectives of the present invention are: to provide improved methods for producing products of formula ^ and other related compounds in high yields; This method of separation steps; and provides such methods that can be implemented with reasonable capital consumption and operated at reasonable conversion costs. Therefore, the present invention is directed to a series of synthetic plans related to Epoxy Rex; Intermediates used in the production of Epileur Reynolds; and the synthesis of such & intermediates. In the description of the preferred embodiment, a novel synthesis scheme is described in detail. The novel intermediate in the present invention Among them, it will be immediately in the following ^ This paper size Gu quiet_ 县 （CNS) # 丨 | (Read the precautions on the back before filling this page)

1T 593305 ΑΊΒ7 V. Description of the invention (3) Those mentioned. The compound of formula IV conforms to the following structure:

IV where -Α · Α- represents a group _CHR4_CHR5 · or _Cr4 = cr5_ R3, R4, and R5 are respectively selected from the group consisting of hydrogen, hydrogen, ceryl, low-carbon alkyl, low-carbon alkoxy, and hydroxyalkane Group, alkoxyalkyl, hydroxycarbonyl 'cyano, aryloxy' R1 represents a low-carbon alkoxycarbonyl or hydroxycarbonyl group, R2 is an 11-releasing group, and its removal is effective at 9_ * u_ There is a double bond between the carbon atoms; -BB-represents the group -CHR6_CHR7- or the group from-or orientation: (Please read the notes on the back before filling in this page j,-口 Γ Staff of the Central Standards Bureau of the Ministry of Economic Affairs Printed by Consumer Cooperatives VCH- -CH — C Hj • CH I • CH-

III wherein R6 and R7 are selected from the group consisting of hydrogen, halogen, low-carbon alkoxy, fluorenyl, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxycarbonyl, fluorenylalkyl, cyano, Aryloxy, and R8 and R9 are selected from the group consisting of hydrogen, halogen, low carbon number oxygen, fluorenyl, 6- This paper size is applicable to China National Standard (CNS) A4 specifications (210X297 mm) 593305 A7 B7 V. Description of the Invention (4-Hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxycarbonyl, alkoxyalkyl, cyano, aryloxy, or ... containing a carbocyclic or heterocyclic ring Structure, or R8 or… together with… or feet 7-carbocyclic or heterocyclic structure 'fused into five per Γ) ring. The compound of formula IV A is equivalent to the compound of formula IV in which R8 and R9 are attached together with a ring carbon to form the following structure: χ, π • A ·]

XXXIV where X, Y1, Y2 and C (l7) are as defined above. The compound of formula IVB is equivalent to the compound of formula IVA in which R8 and R9 together form a structure of χχχιπ: Λ XXXII The compounds of formula IVC, IVD, and IVE printed by the Industrial Cooperative, Ministry of Standards, and Consumer Cooperatives, respectively, are equivalent to each of -A_ A- and -BB. Are -CH2-CH2-, R3 is hydrogen and R1 is an alkoxycarbonyl group, preferably any of formula IV, IV A or IV B of methoxycarbonyl. Compounds in the formula IV range can be prepared by reacting a low-carbon alkylsulfonyl or sulfonating reagent, or a reagent that generates a halide with a corresponding compound in the range of formula V . The compound of formula V conforms to the following structure: This paper size applies to Chinese National Standard (CNS) A4 (210X297 mm) (Please read the precautions on the back before filling this page)

593305 A7 B7 V. Description of the invention (

V wherein -A_A-, -B-B-, R1, R3, R > R9 are as defined in the formula ... The compound of formula V A is equivalent to the compound of formula V in which R8 and R9 are attached together with a ring carbon to form the following structure: (Please read the precautions on the back before filling this page) \ // \ 17 >

CH τ j) t

XXXIV where X, Y1, γ2, and C (l7) are as defined above. The compound of formula VB is equivalent to the compound of formula VA in which R8 and R9 together form a structure of formula χχχιπ: Λ Λ Printed by the staff consumer cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

XXXIII Compounds of formula VC, VD and VE are equivalent to each of -AA- and · Β · Β · are -CH2-CH2-, R3 is hydrogen and R1 is alkoxycarbonyl, preferably methoxyl Of any formula V, VA or VB. Compounds in the range of formula V can be prepared by reacting a metal alkoxide with a corresponding compound of formula VI. The compound of formula VI conforms to the following structure: This paper is suitable for financial use _ Jia Zhuan (CNS) M specification (2iGX297 public attack) 593305 A7 B7

〇 5. Description of the invention (6

VI where _Α-Α-, -Β-Β-, R3, R8, and R9 are as defined in SIV. The compound of formula VIA is equivalent to the compound of formula VI in which R8 and R9 are attached together with a ring carbon to form the following structure: γ, \ / f * / \ ίΤ)

XXXIV where X′Υ1, Υ2 and C (17) are as defined above. A compound of formula vIB is equivalent to a compound of formula VIA in which ... and a structure of formula χχχιπ are formed together: 〇Λ XXXI compounds of formulas VIC, VID, and VIE are equivalent to each of -AA- and -BB- -CH2-CH2- and any of Formula VI, via * ViB, where R3 is hydrogen. Formulas V !, VIA, VIB and V! C are prepared by hydrolyzing compounds corresponding to formula νπ, VIIA, VIIB or VIIC, respectively. The compound of formula VII conforms to the following structure: This paper size applies to the Chinese National Standard (CNS) A4 specification ((Read the precautions on the back before filling this page)

Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 593305 A7 B7 V. Description of Invention (

VII wherein -A_A, -B-B-, R3, R8 and R9 are as defined in formula IV. The compound of formula VIIA is equivalent to the compound of formula VII in which R8 and R9 are attached to the ring carbon to form the following structure: (Please read the precautions on the back before filling in this page)

(CH

XXXIV where X, γΐ, γ2 and C (17) are as defined above. The compound of formula VIIB is equivalent to the compound of formula VIIA in which R8 and R9 together form the structure of formula XXXIII: printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs, ΛΛXXX compounds of formula VIIC, VIID and VIIE, which are equivalent to them respectively Each -AA, -BB- is -CH2-CH2-, and R3 is any of formula VII, VIIA or VIIB, respectively. Compounds in the range of formula VII can be converted by the formula VIII-10- This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) 593305 A7 B7 V. Description of the invention (8) Cyanation to prepare it. The compound of formula VIII conforms to the following structure:

Wherein -A-A, -B-B-, R3, R8, and R9 are as defined in Formula IV.

VIII A compound of formula VIIIA is equivalent to a compound of formula VIII in which R8 and R9 are attached together as a ring carbon to form the following structure:

γ \ / I meaning 17 > 丨 Iim (ch2) 2 · 〇_χ

\ XXXIV where X, Υ1, Υ2, and C (17) are as defined above. The compound of formula VIIIB corresponds to the compound of formula VIIIA in which R8 and R9 together form the structure of formula XXXIII:

(Please read the notes on the back before filling this page)

XXXIII The compounds of formulas VIIIC, VIIID, and VIIIE printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs are equivalent to any formula VIII, where each of -AA- and -BB- are -CH2-CH2, and R3 is hydrogen, respectively. VIIIA or VIIIB. Compounds in the range of formula VIII can be prepared by oxidizing the enzymatic substance including the compound of formula XXX as described below, and efficiently introducing an 11- # group into the enzymatic substance in an α-position via fermentation. Of it. -11 This paper size applies to Chinese National Standard (CNS) A4 specification (210 × 297 mm) 593305 A7 B7 5. Description of the invention (9 The compound of formula XIV conforms to the following structure:

AXIV wherein -A-A-, -B-B-, R3, R8 and R9 are as defined in Formula IV. The compound of formula XIVA is equivalent to the compound of formula XIV in which R8 and R9 are attached together with a ring carbon to form the following structure:

I Υ \ / ί C (1 7) mill (CH2) z. O-x

/ \ XXXIV where X, Y1, Y2, and C (17) are as defined above. The compound of formula XIVB is equivalent to the compound of formula XIVA in which R8 and R9 are attached together with a ring carbon to form a structure of formula XXXIII: Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

(Please read the notes on the back before filling this page)

XXXIII The compounds of the formulae XIVC, XIVD and XIVE are equivalent to any of the formulae XIV, XIVA or XIVB in which each of -A-A- and -B-B is -CH2-CH2_ and R3 is hydrogen, respectively. Compounds in the range of formula XIV can be prepared by hydrolyzing the corresponding compounds in the range of formula XV. -12- This paper size applies to China National Standard (CNS) A4 (210X 297 mm) 593305 A7 B7

V. Description of the invention (1G compound of formula XV conforms to the following structure

AXV • where -B-B-, R3, R8, and R9 are as defined in the formula ... A compound of formula XVA is equivalent to a compound of formula XV in which R8 and R9 are attached together with a ring carbon to form the following structure: (Please read the precautions on the back before filling this page) \ / C (1 7) mill (ch2 ) 2. ό-χ

XXXIV 、 -port Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs

XXXIII where X, Υ1, Υ2 and c (17) are as defined above. A compound of formula XVB is equivalent to a compound of formula XVA in which R8 and R9 are attached together with a ring carbon to form a structure of formula XXXIII: 〇Λ ', πιι / Compounds of formula XVC, XVD, and XVE, respectively corresponding to each of them ~ A and -BB · are any of the formulas χν, χνΑ XVB which are -CHyCH2- and R3 is 氲, respectively. Compounds in the range of formula XV can be prepared by cyanation of the corresponding compounds in the range of formula χνι. -13- + Paper size applies to Chinese National Standard (CNS) A4 specifications (593305 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs A7 _B7 V. Description of the invention (11) The compound of formula XXI conforms to the following structure:

XXI where _A-A-, -B-B_, R3, R8, and R9 are as defined in Formula IV. The compound of formula XXIA is equivalent to the compound of formula XXI in which R8 and R9 are attached together with a ring carbon to form the following structure: νι «Y \ / r / C ^ t7) ,, M, * (CH2) 2 .〇- x xxxiv where X, Y1, Y2, and c (17) are as defined above. A compound of formula XXIB is equivalent to a compound of formula XXIA where R8 and R9 together form a formula:

Q

The compounds of the formulae XXIC, XXID and XXIXE respectively correspond to any of the formulas χχι, XXIA or XXIB in which each of -A-ΑϋΒ_ is -Ch2-Ch2 • and R3 is hydrogen. Compounds in the range of formula xxm can be prepared by hydrolyzing the corresponding compounds in the range of formula X X11. -14- The national standard (CNS) A4 size (2I0X 297 mm) of this paper size applies (Please read the precautions on the back before filling this page)

593305 A7 B7 5. Description of the invention (12 compounds of formula X X Π conform to the following structure:

XXII where -AA-, -B_B_, R3, R8, and R9 are as defined in Formula IV. A compound of formula XXIIA is equivalent to a compound of formula X X11 in which R8 and R9 are attached together with a ring carbon to form the following structure: r \ / IC (1 7) mill (CH “复 · ό_)

XXXIV where X′Υ1, Υ2 and C (17) are as defined above. The compound of formula XXIIB is equivalent to the compound of formula XXIIΑ in which R8 and R9 form a structure of formula χχχιη: (Please read the precautions on the back before filling out this page) Printed by the Consumers Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs Λ \ im / XXX 工Compounds of the formulae XXIIC, XXIID and XXIIE are equivalent to any of formula XXII, XXIIA or XXIIB in which each -AA- and -B · B- are -CH2-CH2- and R3 is hydrogen, respectively. Compounds in the range of formula XXII can be prepared by cyanation of corresponding compounds in the range of formula XXIII. -15- The dimensions of this paper are applicable to Chinese National Standard (CNS) A4 (210X297 mm) 593305 A7 B7 V. Description of the invention (13 The compound of formula XXIII conforms to the following structure:

XXIII where -B-B_, R3, R8 and R9 are as defined in SIV. 1 The compound of formula XXIIIA is equivalent to the compound of formula X X111 in which R8 and R9 are attached together with a ring carbon to form the following structure: \ / Π mm (ch2)

XXXIV where X, Y1, Y2 and C (17) are as defined above. The compound of formula XXIIIB is equivalent to the compound of formula XXIIIA where R8 and R9 together form a structure of formula χχχπι:

(Please read the notes on the back before filling this page}

It is required to print compounds of formula XXXIII of formula XXIIIC, XXIIID, and XXIIIE by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs, which are equivalent to any formula XXIII in which each -AA- and -BB · are -CH2-CH2- and R3 is hydrogen XXIIIA or XXIIIB. Compounds in the range of formula XXIII can be prepared by oxidation of the corresponding compounds in the range of formula XXIV, as described below. The compound of formula 104 conforms to the following structure: -16- This paper size applies to the Chinese National Standard (CNS) A4 specification (210X297 Gongchu) 593305 A7 B7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (14 〇

104 where _A_A_, _B_B_, and R3 are as defined in the formula, and r11 is Ci to q alkyl. The compound of formula 104A 'is equivalent to a compound of formula 104 in which each of _a_a _ * _ b_b_ is _CH ^ CH2- and R3 is chloro. Compounds in the range of Formula 104 can be prepared by thermal decomposition of compounds in Formula 103. The compound of formula 103 has the following structure:

CO <103> wherein -A-A-, -B-B ·, and R3 and R11 are as defined in Formula 104. The compound of formula 103A is equivalent to the compound of formula 103 in which each of _a_a _ * _ b_b_ is -CI ^ -CH2- and R3 is hydrogen. For compounds in the range of Formula 103, the corresponding compound of Formula 102 and dialkyl malonate can be used (Please read the precautions on the back before filling this page)

-17- 593305 R3

Where -Am, R3 and RU are as defined in Formula 104. 〇

Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7 B7 V. Description of the Invention (IS) The compound of formula 102 prepared by the reaction in the presence of osmium oxide, such as a gold test compound, conforms to the following structure: 102 Compound of formula 102A, Corresponding to a compound of formula 102 in which each of _a_a _ * _ b_b_ is -CH ^ CH2- and R3 is hydrogen. A compound in the range of Formula 102 can be prepared by reacting a corresponding compound of Formula 101 with a trialkylphosphonium in the presence of a base. The compound of the formula 101 corresponds to the following structure: 101 where Α-Α_, Β-Β-, R3 and R11 are as defined in the formula 104. The compound of formula 101A is equivalent to the compound of formula 10 in which each of -α_α * _β_β_ is CH ^ CH2- and R3 is hydrogen. The valence compound in the formula 10 can be reacted with trialkyl orthoformate in the presence of an acid by means of 1 1 "hydroxyandrostene-3,17_dione or other ^ XXXVI compounds ^ Be prepared. ~ -18- This paper size applies Chinese National Standard (CNS) Α4 size (21〇χ 297 mm) (Please read the precautions on the back before filling this page)

593305 A7 B7 5. Description of the invention (16) The compound of formula XL conforms to the following structure:

90

XL where -E-E- is selected from the following, R "| XL workers (please read the precautions on the back before filling this page): c-c-

XLIV · 〆4, βμΙ C— c

XLV ONIC, printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs

: lc—R XLVI y and XLV workers 19- This paper size is applicable to Chinese National Standard (CNS) A4 (210X 297 mm) 593305 A7 B7 Five invention descriptions (17 Han "is ... to ^ alkyl, or Rl8 〇_Group—from the formation of the _environmental bridge, R ", R" and R "are selected from the group consisting of hydrogen, alkynyl, and sulfonamide: and cyano 'R24it from hydrogen and low carbon number alkynyl' r8> r9 () Are selected from di and include R and R (as defined above, with reference to formula ...), and substituents -AA-, -BB-, R3 as defined in formula 104, respectively. 'Compounds of formula XL A Equivalent to formula XL in which r2i, r22 and r23 are selected from the group consisting of gas, halogen, and low-carbon alkyl, respectively. Compounds of formula XLB are equivalent to _E_E_ which conform to formula XLU,, and 1 ^ ¥ or 1 ^ ¥ 11 The compound of the formula 乂 1 ^ 八. The compound of the formula 1 又 is equivalent to the formula XLB where _E-E_ conforms to the formula XLV. The compound of the formula XLD corresponds to the formula XLB where EE- complies with the formula XLVII. The compound of the formula XIE is equivalent In which r8Q and r9❶ are together with the ring carbon atom attached thereto, including a ketone or: \ /, I 7) mm (ch8) ^. 〇m)

XXXIV (Please read the notes on the back before filling out this page) • For example, 11 The Consumer Cooperatives' Seal of the Central Standards Bureau of the Ministry of Economic Affairs f where X′Υ1, γ2, and c (17) are the same as defined above. XLF, XLG, The compounds of XLH, XLJ, XLM and XLN are equivalent to _ AA- and -BB- and R3, respectively, as defined above. The compounds of formula xl, XLA, XLB, XLC, XLD and XLE. The compounds of formula XL I conform to the following structure. :

(See plan 9) XLI This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) V. Description of the invention (I8 where -EE- is selected from the group consisting of: R * s, Re, | 0 * — C-: C-c— t1〇R24, Rm | C—Q—, u OR \ lc ~ 24 A7 B7 xLXli XLiv

XLV and

• L XLVi: cfibR- XLVI i Printed by the Consumers' Cooperative of the Central Government Bureau of the Ministry of Economic Affairs

R18 is Cl to C4 alkyl, or R18O &amp; base bridge, R21, R22 and r23 are selected from hydrogen, minus, halogen, and radical, respectively; r24 is selected from hydrogen and low carbon number alkyl, r80 and r9G And a substituent selected from and including R and R 9 (as defined above, with reference to Formula IV), and -AA-, -B-B_, and R3 as defined in 104, respectively. The compound of the formula XLIA is equivalent to the formula XLI in which R22 and R23 are selected from the group consisting of halogen, and a lower alkyl group, respectively. The compound of the formula XLIB corresponds to the formula XLIA in which the formula XLIn, XXL or XLVII is met. The compound of XLIC is equivalent to that in which r9 is formed together with the group attached to it.

(Please read the notes on the back before filling this page)

593305 A7 B7 V. Description of the invention (I9) Carbon atoms together, containing ketones or the following formula XL 1: \ /] 7) mu, (〇h8) f.

XXXIV

X, Y1, Y2, and C (l7) are as defined above. The compound of the formula XL ID is equivalent to the formula XLID r \ / f / <in mm (ch2) 2 in which the substituent χχχΙν conforms to the structure XXXIII

XXXIV Read extensively first and read the back 1 Note that the compounds of formula XL IE that are really filled in are equivalent to formula XL in which R80 and R90 together form a ketone. The compounds of the formulas XLIF, XLIG, XLIH, XLIJ, xlim and XLIN are equivalent to the formulas XLI, XLIA, XLIB, XLIC, xlid and XLIE, respectively, where -A-A_,-and R3 are as defined above. Compounds in the range of formula XLI can be prepared by hydrolysis of the corresponding compounds of formula B and B as defined below. The compound of formula XLII conforms to the following formula:-Ordered by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

.η • 0 XL · Gonggong -22- The paper size applies to Chinese National Standard (CNS) A4 (21 OX 297 mm) 593305 A7 B7 V. Description of the invention (20 c-c-: C-c-

XLIII XLIV if -c— 24

XLV OR24, i— R41

Printed by XLVI and the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs

f XLVII R21, R "and R23 are selected from hydrogen, alkyl, halo, nitro and cyano, Rj4 is selected from hydrogen and low-carbon alkyl, R80 * R9G is selected from ketone and may include R and R And Α-Α-, -Β-Β_, Ι13 are the same as the substituents defined in formula ι04. The compound of formula XLIIA is equivalent to wherein R2 !, 22 and 23 are selected from hydrogen, halogen and A low-carbon alkyl group of the formula XL jj. A compound of the formula XLIIB corresponds to the formula XLIIA where -E · E_ conforms to the formula XLni, XLIV, XLV or XLVII. &Lt; A compound of the formula XLIIC corresponds to where r80 and r90 are equal to The ring carbon atom attached to it contains a ketone or the following formula XLII: γι Qing Y \ / τmm (che) t XXXIV 23- This paper size applies to the Chinese National Standard (CNS) A4 specification (21 OX 297) Li) (Please read the notes on the back before filling this page)

593305 A7 B7 V. Description of the invention (21 where X'Y1, Y2, and C (17) are as defined above. The compound of formula XL I ID is equivalent to the formula in which the substituent XX XIV conforms to the structure XXXIII XLIID 〇

The compound of the XXXII formula XLIIE is equivalent to the formula XLIID in which R80 and r9G together form a ketone. The compounds of the formulas XLIIF, XLIIG, XLIIH, XLIIJ, XLIIM and XLIIN are respectively equivalent to the formulas XLII, XLIIA, XLIIB, XLIIC, XLIID and XLIIE where _Α_ΑϋΒ_ is _CH2_ CH2, respectively, and is hydrogen. Compounds in the range of formula XLII can be prepared by deprotecting the corresponding compounds of formula X LI. The compound of formula XLIX conforms to the following structure:

R ·

XLIX (Read the precautions on the back before filling this page) _ ^.

, 1T Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs where -E-E- is defined as in Formula XL, and _a_a_, _b_b., R1, R3, R8, and R9 are defined as in Formula V. The compound of formula XLIXA is equivalent to the formula XLIX in which the ring carbons attached thereto form the following structure together: -E-E · \ ^ CM7) -24-

XXXIV 593305 A7 B7 V. Description of the invention (22 where X, Y, Y2 and C (17) are as defined above. The compound of formula XLIXB is equivalent to the formula XLIXA where r8 and r9 together form the structure of formula XXXIII: Λ 'mu / XXX Compounds of the formula XLIXC, XLIXD, XLIXE are equivalent to any of the formulas XLIX, xlixa or XLIX, xlixa or XLIXB. Compounds in the range of formula XLIX can be prepared by reacting an alcohol-containing or water-containing solvent with the corresponding compound of formula VI in the presence of a suitable base. The compound of formula A203 has the following structure:

[A203] (Please read the precautions on the back before filling out this page}. · · · Orders selected from the following: Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

• R ”, RI1I C—Q. / II XLIII R21 XLIV 25 This paper size applies Chinese National Standard (CNS) A4 specification (210X297 mm) 593305 A7 B7 V. Description of the invention (23 • 1〇R, R * 1 | c ~ *

XLV OR24 Rai% —i— / I »

XLVI and

XLVI I Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs R18 is selected from C! To C4 alkyl groups, R21, feet 22 and feet 23 are selected from hydrogen, alkyl, halogen, nitro and cyano, and R24 is selected from hydrogen and Low carbon number alkyl, and -BB-, R3 have the same meanings as defined in Formula 104. The compound of formula A203A is equivalent to formula A203 in which r2i, feet 22, and feet 23 are selected from hydrogen, halogen, and lower carbon alkyl, respectively. The compound of formula A203B is equivalent to formula A203A in which _E_E_ conforms to formula XLIII, XLIV, XLV or XLVII. The compounds of the formula A203C, A203D and A203E respectively correspond to the formula A203, A203A or 203B in which each of -A-A and -B-B- is -CH2-CH2-, and R3 is hydrogen. Compounds in the range of formula A203 can be prepared by careful reduction of compounds of formula A202 as defined below. The compound of formula A 2 0 4 conforms to the following structure:

[A204] (Please read the notes on the back before filling in this page}-,-=-口-Γ This paper size is applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) 593305 type A204C, A204D and A204E Compounds corresponding to formulas A204, A204A, and A204B in which each -AA- and -BB- are -CH2-CH2-, and r3 is hydrogen. Compounds in the range of formula A204 can be obtained by formula A It is prepared by hydrolysis of the corresponding compound of 203. The compound of formula A 205 conforms to the following structure:

Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7 B7 V. Description of the invention (24 of which R19 is 1 to (: 4 alkyl), and _E_E_, _a_a_, _b_b_ * r3 are as defined in Formula 203. Formula A2 The compound of 04A is equivalent to the formula A2 04 in which R22 * R23 is selected from hydrogen, halogen and low-carbon alkyl, respectively. The compound of formula A204B is equivalent to the formula A204A in which _E_E conforms to the formulas Xlm, xuv, XLV or XLVII. [A205] wherein R 0 is (^ to stilbyl) and-, R19, · A-A-, -B_b &gt; ^ r3 are as defined in formula 203. A compound of formula A205 A is equivalent to r2i, r22 and r23 are respectively selected from the group consisting of hydrogen, halogen and low-carbon alkyl formula A205. The compound of formula A205B is equivalent to formula A205A in which _E_E_ conforms to formula XLIII, XLIV, XLV or formula XLVII. Formulas A205C, A205D and The compounds of A20 5E are equivalent to each of -AA- and -BB, which are &lt; η2-(: Η2 ..., and R3 is hydrogen, respectively. -27 · This paper size applies Chinese National Standard (CNS) A4 specifications ( 210X 297 mm) (Please read the notes on the back before filling in this page j

593305 A7 B7 5. Description of the invention (25) A205, A205A and A205B. Compounds in the range of Formula A205 are prepared by reacting the corresponding compound of Formula A204 with an alkanol and an acid. The compound of formula A 2 0 6 conforms to the following structure: may 2 0

[A206] B-B-and R3 are the same as those printed in 205 in the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs, where R19 and R are defined. The compound of the formula A206A is equivalent to the formula A206 in which R21, R22 and R23 are selected from hydrogen, halogen and a low carbon number group, respectively. The compound of formula eight 2066 is equivalent to the formula A206A in which-£-£-conforms to formula 1 ^ 111, XLIV, XLV or formula XLVII. Compounds of formulas A206C, A206D and A206E, respectively, J are equivalent to formulas A206, A206A and A206B in which each of -A-A- and -B-B- is -CH2-CH2 · and R3 is hydrogen. A compound in the range of Formula A206 can be prepared by reacting a corresponding compound in the range of Formula A205 with a trialkylphosphonium halide. The compound of formula A2 0 7 conforms to the following structure:

COffR2i [A207] f Please read the notes on the back before filling in this page j 1 «· -Order · This paper size applies to China National Standard (CNS) A4 specification (210X297 mm) 593305 A7 B7

Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs. 5. Description of the invention (26 where R25 is (^ to! ^ alkyl, and -EE-, R19, R20, -AA-, and R3 are as defined in Formula A205). The compound of formula A207A is equivalent to the formula A207 in which r2i, R22 and R23 are selected from hydrogen, halogen and a low-carbon alkyl group, respectively. The compound of formula A207B is equivalent to -EE- in accordance with formula XLIII, XLIV, XLV or XLVII. Formula A207A. Compounds of Formula A207C, A207D, and A207E are equivalent to Formula A207, A207A, and A207B, each of which -AA- and -BB are _CH2-CH2-, and R3 is hydrogen. Within the scope of Formula A207. The compound of formula A 2 0 8 can be prepared by reacting the corresponding compound of formula A 2 0 6 with dialkyl malonate. The compound of formula A 2 0 8 has the following structure [A208] where -EE-, R80 and R% are as in The formula is as defined in £ 11, and _A_A_ BB- and R3 are as defined in formula 104. r19, r20, _octadecane, and R3 are as defined in formula 205. The compound of formula A20 8A is equivalent In which R2i * R22 is selected from the group consisting of hydrogen halide and low-carbon alkyl, respectively, formula A208. The compound of formula A20 8B is equivalent to _EE _Compliant XLln, (Please read the precautions on the back before filling this hundred c)

29- 593305 A7 B7

Q

V. Description of the invention (27 XLV or formula A208A of formula XLVII. The compound of formula A208C is equivalent to R8G and r9G, together with the ring carbon attached to it, containing a ketone or formula A20 8 of the formula:

XXXIV where X′Υ1, Υ2 and C (17) are as defined above. The compound of formula A208D is equivalent to formula A2 0 8D in which the substituent χχχιν conforms to the structure of formula XXXIII:

XXXIII Compounds of the formula A208E, A208F, A20 8G, A208H and A208J, respectively equivalent to the formulas A208, A208A, A208B, A208C, or _AA- and -BB-, respectively, and R3 is hydrogen A208D. Compounds in the range of formula A208 can be prepared by thermal decomposition of the corresponding compounds of formula A2 07. The compound of formula A2 0 9 conforms to the following structure: (Please read the precautions on the back before filling this page}

Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs, ΦΟ

R · [A209] where 118 () and 119 () are the same as in the formula; ^ 1, and 44-, -eight-eight-, -30- This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 (Mm) 593305 A7 B7 5. Description of the Invention (28) -BB- and R3 are as defined in Formula 205. The compound of the formula A2 09 A is equivalent to the formula A209 in which R21 and R22 are selected from the group consisting of Li, halogen, and lower alkyl. The compound of the formula A2 0 9B is equivalent to the formula A209A in which -E-E · conforms to the formula XLIII, XLIV, XLV or XLVII. The compound of formula A209C is equivalent to -EE-complying with formula A209B of formula XLIV. 0 The compound of formula A209D is equivalent to where R8G and r9g, together with the ring carbon attached thereto, contain a ketone or formula A2 09: Y \ / F / C \ 7) ,,,, N -0-Χ

XXXIV where X, Υ1, γ2, and c (17) are as defined above. The compound of formula A209E is equivalent to the formula 8209 in which the substituent χχχΙν conforms to the structure of formula 111.

xxici Ministry of Industry, Economy, Trade and Industry, Central Standards Bureau, Employees' Consumer Cooperatives, India (please read the precautions on the back before filling this page) The compounds of formula A209F, A209G, A209H, A209J, A209L and A209M are equivalent to each of them -A_A- And formulas A209, A209A, A209B, A209C, A209D, and A209E, which are _Ch2_CH2- and R3 is hydrogen, respectively. Compounds in the range of formula A209 can be prepared by hydrolysis of the corresponding compound of formula A208. The compound of formula A 2 1 0 conforms to the following structure: -31- The paper size is suitable for financial and family care; f bound (CNS) A specification (210X297 gong) 593305 A7 B7 V. Description of the invention (29 R η · 〇

[A210] wherein R80 and R90 are as defined in Formula XLI, and the substituents _α_α_, B-, and R3 are as defined in Formula 104. The compound of formula A210A is equivalent to the formula A2 10 which contains ketone or r9〇 and the ring carbon attached to it: Β- (Please read the precautions on the back before filling this page) 丨 丨 丨丨 丨 丨

XXXIV Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs Λ 'iim / where X, Y1, Y2, and C (17) are as defined above. The compound of the formula A2 1 0B corresponds to the formula A210A in which the substituent χχχιV conforms to the structure of the formula XXXIII: XXXIII _ The compound of the formula A2 10C corresponds to the formula A210A in which ketone is formed together with r90. Compounds of formula A2 1 0D, A210E, A210F, and A210G, respectively, are eliminated by each of the formulas -32- 593305 A7 B7 where -AA- and -BB- are -CH ^ CH2 · and R3 is hydrogen (30A210, A21〇a, A210B, and A210C. Compounds in the range of Formula A210 can be prepared by the epoxidation of a compound of Formula 209 where -E_E • is> C-CH_. Formula A2 1 1 The compound conforms to the following structure:

fT

[A211] wherein -A-A-, _Β_Β · and R3 are as defined above. The compound of the formula A211Α is equivalent to the rule wherein the ruler 80 and the ruler 9 together contain a ketone or the formula: A 2 1 1: r \ / J 17) min

XXXIV (Please read the precautions on the back before filling this page} Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs, where χ, γΐ, Y2, and c (17) are as defined above. The compound of formula A2 1 1B is equivalent to it The substituent χχχιν conforms to the formula A211A of the structure of formula XXXIII:

XXXIII Compounds of formula A2 1 1C are equivalent to form together with r90. They are the same as A210A 0-33 593305 A7 B7 V. Description of the invention (31) Compounds of formula A211D, A211E, A211F and A211G are equivalent to each of -AA -And -BB- are the formulas A211, A211A, A211B, and A211C, each of which is -CHrCHr and R3 is hydrogen. Compounds in the range of formula A211 can be oxidized by the corresponding compound of A210 or in the epoxidation of the corresponding compound of formula A209 where ^^ is &gt; ^-crM 'as described below It is converted into a compound of formula 11 to prepare it. The compound of formula L conforms to the following structure: (Please read the notes on the back before filling this page}

Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economics where R is Ci to C4 Pit ’and -A-A-, -B-B_, R1, R2, r3, and R9 are as defined above. The compound of formula L A is equivalent to formula L where R8 and R9 are together with the carbon atom attached to it, including: YI t γ \ / f / C \ 7} Π ,, Π ί -ό ^ χ

XXXIV where X'Y1 and Y2 are as defined above.

The compound of formula LB is equivalent to formula L where R8 and R9 conform to formula.

593305 A7 ------ B7_ V. Description of the invention (32) '&quot; Compounds of the formula LC, LD, and LE are equivalent to · A-Α · and-& B- are each -CHrCH2-, and R3 is a formula L, LA and LB of hydrogen. (Please read the notes on the back before filling out this page) Based on the disclosure of a specific reaction plan as stated below ’These compounds that are most useful for a particular reaction plan will be obvious. The use of the compound of the present invention can be used as an intermediate of epoxy mexrenol and other steroids. Other goals and characteristics are partly obvious and part will be pointed out later. Description of the M film. Figure 1 is a flow chart of the plan for the biological conversion method of canrenone or a Conreno derivative into a relative 1 1 α-hydroxy compound. Figure 2 is the -hydroxy group of a Conreno or Conreno derivative. Figure 3 is a flow chart of the best method for biological conversion of chemical conversion; Figure 3 is a flow chart of the best method for biological conversion of 1 1 _ α-light-based light conversion of Conrad or Conreno derivatives; The specific size distribution of Conorno prepared by the method of 2; and FIG. 5 shows the specific size distribution of Conorno sterilized in the conversion fermenter according to the method of FIG. 3. Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. The corresponding reference characteristics indicate the corresponding parts in the entire picture. Description of the preferred embodiments According to the present invention, in order to prepare epoxy mexanol and other compounds conforming to the formula, various novel method plans have been devised: -35- This paper size applies Chinese National Standard (CNS) Α4 Specifications (210 × 297 mm) called 05 A7 B7 Five 'invention description (33

Where: • AA- represents the group _CHR4-CHR5- or -CR4 = CR5- R, R4 and R5 are respectively selected from the group consisting of hydrogen, halogen, borrowed group, low carbon number fluorenyl group, low carbon number alkoxy group, hydroxyalkane Group, alkoxyalkyl group, hydroxycarbonyl group, cyano group, aryloxy group, R1 represents α-azimuth low-carbon alkoxycarbonyl or hydroxyalkyl group, and represents the group _CHr6-cHR7- or from or /? _ Azimuth group; CH- I -CH- CH- -in.

Industrial and Industrial I, Consumer Standards Cooperative of the Central Bureau of Standards, Ministry of Economic Affairs, where R6 and R7 are respectively selected from the group consisting of hydrogen, halogen, low-carbon alkoxy, fluorenyl, triphenyl, alkoxyalkyl, hydroxycarbonyl, alkyl, Alkoxycarbonyl, fluorenyl, cyano, aryloxy, and R8 and R9 are selected from the group consisting of hydrogen, hydrogen, low-carbon alkoxy, fluorenyl, benzyl, oxyalkyl, and Group, alkyl, alkoxycarbonyl, alkoxycarbonyl, cyano, aryloxy, or R8 * R9 together contains a carbocyclic or heterocyclic structure 'or R8 or R9 together with R64R7 contains a carbocyclic or The heterocyclic structure is fused into a five-ring D ring. Unless otherwise stated, in this disclosure, "low carbon number" is specified as -36- This paper size is applicable _ house standard (CNS) A4 specification (21GX297 mm (please read the precautions on the back before filling in (This page)

593305 A7 B7 V. Description of the invention (34 The organic group contains a maximum of 7, preferably from 1 to 4 carbon atoms. The low-carbon-fe oxycarbonyl group is preferably derived from a morpho group having 1 to 4 carbon atoms. 'Like methyl, ethyl, propyl, isopropyl, butyl, isobutyl, first-butyl and second-butyl; particularly preferred are methoxycarbonyl, ethoxyfluorenyl and isopropyl Propoxycarbonyl. The low carbon number alkoxy group is preferably derived from the c 1-c 4 alkyl group mentioned above, especially derived from the primary alkyl group; particularly preferred is methoxy group. Low carbon number chain Carbofluorenyl is preferably derived from a linear alkyl group having from 1 to 7 carbon atoms; most preferred are methylfluorenyl and ethylfluorenyl. Methylene bridges between the 15, 16, 6-positions, most It is 0-azimuth. According to the method of the present invention, the preferred compounds are the 20-spiroxane compounds described in U.S. Patent No. 4,559,332, which are those that conform to formula IA: (please first Read the notes on the back and fill out this page} _t ·, -Order

Printed by the Staff Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs

• c—X ΙΑ where:-Α_Α- represents the group _CH2-Ch2kh = ch_, the table group _CH2_CH2- or the group of the formula IIIA or the 々-position group 'Η 一-C Η- —CH- R1 represents ^ -Position of low-carbon alkoxycarbonyl or hydroxycarbonyl groups, χ represents two hydrogen atoms, oxo or = s, -37 _ This paper size is applicable to Zhongguanjia Standard (CNS) M specification (mu97 公 酱) 工 工 ΙΑ Staff Consumer Cooperatives, Central Bureau of Standards, Ministry of Economic Affairs

593305 A7 --- —__ B7 V. Description of the invention (35) Y1 and γ2 together represent the oxygen bridge _〇_, or γ 1 represents the # 1 group, and Υ2 represents a hydroxyl group, a low-carbon alkoxy group, or if χ On behalf of η2, it may also be a low-carbon-chain alkoxy group. The 20-spirooxo compounds produced by the novel method of the present invention are preferably those of formula J in which Υ1 and Υ2 together represent the oxygen bridge _〇_. Particularly preferred compounds of formula I are those in which X represents oxo. Among the compounds of the formula IAi20.spiro oxycarbons in which X represents oxo, the most preferred are those in which Y1 and Y2 together represent the oxygen bridge _〇_. As already mentioned, 17y5_hydroxy_21_carboxylic acid can also exist in its salt form. Special consideration is given to metals and ammonium salts, such as metal and soil metal salts, such as steel, iron, copper and better treacherous salts, as well as derivatives derived from ammonia or suitably, preferably, physiologically Tolerable organic-nitrogen-base = ammonium salts. Because amidine is not only considered amines, such as low carbon number alkylamines (such as triethylamine), hydroxy_low carbon number alkylamines [such as 2-hydroxyethylamine, diethyl) amine or tri- ( 2-Byethyl) amine], cycloalkanylamines (such as di% hexylamine) or amines (such as amine and N, N, dimethylethylenediamine), and _ nitrogen-containing Cyclic compounds, such as those with aromatic characteristics (such as elu or quilinyl), or those with at least one saturated heterocyclic ring (such as N_ethylπ, pyridine, morpholine, hexahydro Pycnogenol or N, N, -dimethylhexahydropyridine). Alkali metal salts are also included in the preferred compounds, in particular where R 1 represents fluorenyloxycarbonyl, X represents oxo and γ &gt; 2 represents the unsalting salt of the hydrazone compound of Mystery formula. Particularly preferred compounds of formula I and IAA are, for example, the following: -38- This paper size applies to the Zhongguanjia Standard (CNS) A4 specification (2) Gx297 (Please read the precautions on the back before filling this page)

Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs 593305 Μ —__ Β7 V. Description of the Invention (36) 9 (^, 11 (^-epoxy_7 &lt; ^-methoxyisopropyl-20-spiroxy_4_ 晞 _ 3,21-dione, 9α, 11 from epoxy-7 from -ethoxycarbonyl-20-spiro-4-ene-3,21-dione, 9α, 11 epoxy isopropoxycarbonyl_2 (μ Spiro-4-en-3,21-dione, and 1,2-dehydro analogues of each of the compounds, 9α, 11 Han-epoxy-6 Han, 7 Han-methylene-20-spiro -4 · ene-3,21_diketone, 9α, 11 from -epoxy-6yg methylene-2O-spiro-4_ 晞 _3,21_diketone, 9 α, 11 α -epoxy _6 Lu, 7 Lu; 15 / ?, 16 Lu-bismethylene_2〇_spiro_4 • 晞 _ 3,21 -diisomorphic, similar to the 1,2-dehydrogenation of each of these compounds Compound, 9σ, 11 α · epoxy-7α-methoxycarbonyl-17col-¾yl-3 -oxo-pregnant · 4-ene-2 1-carboxylic acid, 9 ", 11" -epoxy-7 "-Ethoxycarbonyl_17 /?-Hydroxy-3-oxo-progesterone_ 4-ene-21-carboxylic acid, 11" -epoxy-7α · isopropoxycarbonyl-17 lu · hydroxy-3- Oxo-progesterone-4-pyrene- 21-retedic acid, 9α, 11 α-epoxy_17cold-light-based-6 &lt; ^, 7 from-methylene-3-oxo-progesterone · 4_晞 -21_ a few acids, 9 ", η α-epoxy] 7 cold-hydroxy-6 cold, 7 /? _ Methylene-3 · oxo-progesterone-4- cotton · _21-chinic acid, 9 with, 11" mono epoxy -17; 5- # presents the group -6 / ?, 7 ^; 15 / ?, 16y5 -bismethylene-3 -oxo-pregn-4-4ene-2carboxylic acid, and alkali metal salts , Especially the desalted or ammonium of each of these acids, and the corresponding 1,2-dehydrogenated analogs of each of the carboxylic acids mentioned, or their salts. 9α, 11 α -epoxy-i5y5, Uy5_ Methylene · 3,2ΐ_ Dioxo-20-Spiral • 39- This paper size applies to China) Μ size (2 丨 〇 &gt; &lt; 297 public attack) ~ &quot; (Please read the precautions on the back first Fill out this page}

Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs 593305 A7 -------- "5. Description of the Invention (37) Oxypene-7"-methyl carboxylate, ethyl ester and isopropyl ester, -9q, 11o ^ epoxy-15 / ?, 16々_ methylene_3,21_ dioxo · 20_spiro-1,4-diene-7 π-carboxylic acid methyl ester, ethyl ester and isopropyl ester , 9 ", 11" -epoxy-3_oxo_2〇-spiro-4-ene-7 "_formic acid carboxylic acid, ethyl and isopropyl ester, 9 α, 1 1" _ Epoxy-6 cold, 6 /? _ Methylene_2.spiro-4_-3_one, 90%, 11-epoxy-6 Lu, 7 Lu; 15々, 16; 3- Bismethylene-20-spiro-4-ene_3-one, and 9α, 11α_epoxyhydroxy_17_ (3.hydroxy.propyl 3_oxo_androst-4-ene- 7 from methyl carboxylate, ethyl ester, and isopropyl ester, 9 ", 1 1 α _epoxy-17 • hydroxy_17 α _ (3_hydroxypropyl) _6 to 7" _ methylene-androster -4 -fluorene-3 -one, 9 from, 11 α _epoxide_17fluorene_hydroxy-17 α _ (3_hydroxypropyl) · 6 cold, methylene-androster-4 _fluorene-3- Ketone, 9 from, 11 α-J oxo · 17々_hydroxy-17 "_ (3_hydroxypropyl) _6々, 70; 15 points, 16々 · bismethylene · androst_4_ene- 3_ ketone, including extract &lt; 17α- (3-ethoxypropyl) and 17- (3-methyloxypropyl) analogs of androstane compounds, and all of the androstene mentioned 4_ 4-3 · Ketones and 1,2-dehydro analogues of compounds of the 〇_spirooxy_ 4 _ene_ 3 _ ketone series. Chemical names of compounds of the formulae I and IA &lt; and similar compounds having the same characteristic structural type, are According to the current nomenclature, it is derived in the following way: About, Zhongzhong and Υ together represent _〇_, which is derived from 20-spiroxane (for example, where X represents oxo, and γι and the representative formula) This paper size applies the Chinese National Winter Standard (CNS) A4 Regulation (2H) X 297 Meal) (Please read the precautions on the back before filling this page)

593305 A7 B7 V. Description of the invention (38 spiro-oxygenated _21,); about which γ1 and γ2 respectively represent the following: those of generation I are derived from 17 /?-Hydroxy-17 from _pregnant 晞 2 1-carboxyl Acid; and with respect to and by i ^^, /, in which Y and Y respectively represent a hydroxyl group, and X represents two lice: those that are derived from 17 / ?, the group _17 ^ (3_light propyl), Junda &amp; So unless specifically mentioned otherwise, in the final product of Formula I and similar structures, shells and intermediates, in each case, the shapes mentioned above and below are understood because of the ring and opening The form of the chain, that is, the intrinsic intrinsic and 17 /? 'Yl_21 side acids and their salts, are so closely related to each other that the latter can only be regarded as the former hydrated form. According to the present invention, to prepare the compound of formula I at high yields and reasonable costs, several different schemes have been devised. Each synthesis plan is carried out through the preparation of a series of intermediates. Many of these intermediates are novel compounds, and the methods for preparing these intermediates are also novel methods. 3L scoring 1 (starting from Conranox, a better plan for the preparation of phantom compounds, and advantageously starting from Conranox or Phenol X111 related starting materials (please read the precautions on the back before filling in this Page) Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs

XII-AA-representing group -CHR4-CHR5KH4 = CH5_ R3, R4 and R5 are respectively selected from the group consisting of hydrogen, halogen, hydroxyl, low-carbon alkyl and low-carbon oxygen, per-alkyl, oxy-alkyl, Super carboxyl, cyano, aromatic

4 -41- This paper size applies Chinese National Standard (CNS) A4 specification (210X 297 public holidays) 593305 A7 B7 V. Description of the invention (39 oxygen, _b-b_ represents the group-CHR6_CHR7_ or the base of the heart or orientation Mission:,? Η · -CH —CHa

-CH I -CH

IU wherein R6 and R7 are respectively selected from the group consisting of hydrogen, halogen, low-carbon alkoxy, fluorenyl, alkynyl, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxycarbonyl, alkoxyalkyl, cyano, Aryloxy, and R8 and R9 are selected from the group consisting of hydrogen, halogen, lower alkoxy, fluorenyl, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxycarbonyl, alkoxyalkyl, Cyano, aryloxy, or R8 and R9 together contain a carbocyclic or hetero i structure, or R8 or R9 and R6 or R7 together contain a carbocyclic or heterocyclic ring, = structure, fused into a pentacyclic ring D ring. . Using a biological transformation method of the type explained in FIGS. 1 and 2, the 11-hydroxy group of α · square ^ is introduced into a compound of formula XIII, thereby obtaining a compound of formula: ^:

Printed by the Staff Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs

VIII where _A-A-, -B-B-, R3, R8, and R9 are as defined above. The compound of formula XIII has a structure: Y2 H C γ 1 I ⑶—rWH factory

Righteousness. Preferably,

XXXA (Please read the notes on the back before filling in this page j

-42- 593305 A7 B7 V. Description of the invention (40 and the product of 1 1 α-hydroxyl has structure: H.r Y1

X VII ΙΑ where each Α_Α represents a group ί! ΙΗ2_ (ΙΉ2 · or, -Β · Β · represents a group -CH ^ CH2 ·, or ^ or 々. Orientation group: -CH— CHP— CH—-

IIIA X represents two hydrogen atoms, oxo or, Y1 and Y2-together represents an oxygen bridge-0_, or Y1 represents a hydroxyl group, and Y represents per group, a low carbon number alkoxy group, or if X represents h2, it can also be substituted ^ Low-carbon alkanoyloxy, and salts of compounds in which X represents oxo and Y2 represents hydroxyl, "The compound of formula VIII produced in this reaction conforms to formula Viha (please read the notes on the back before filling in (This page) Printed by the Consumers' Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs 〇〇 斗斗 YccH2] "c: =: x

VII ΙΑ -43- This paper size applies to China National Standard (CNS) Α4 specification (210 '乂 297 mm)

Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 593305 A7 B7 V. Description of the Invention (41) Among them -A-A-, -B-B-, Y1, Y2, and X are as defined in the formula XXXA. More preferably, R8 and R9 together form a 2 0 -spiroxane structure:

XXXIII -A-A- and -B-B- are -CH2-CH2-, and R3 is hydrogen. Preferred organisms that can be used in this hydroxylation step include Aspergillus ochraceus NRRL405, Aspergillus ochraceus ATCC 18500, Aspergillus niger ATCC 16888 and ATCC 26693, Aspergillus ni du 1 ans ATCC 1 1 267, Rhizopus orvzae ATCC 11145, Rhizopus stolonifer, ATCC 6227b, Streptomyces fradiae ATCC 10745, Bacillus megaterium ATCC 14945, Pseudomonas cruciviae ATCC 13262, and Trichothecium roseum ATCC 12543. Other preferred organisms include Fusarium oxysporum f sp. cepae, ATCC 11171, and Rhizopus arrhizus ATCC 11145 ° Other organisms that have shown activity against this reaction, including Absidia coerula ATCC 6647, P. gray-green (Absidia glauce) ATCC 22752, Actinomucor elegans ATCC 6476, Aspergillus flavipes ATCC 1030, fumigant Aspergillus fumigatus ATCC 26934, Beauveria bas—ATCC 7159 and ATCC 13 144, Grape Cavity-44- This paper applies Chinese National Standard (CNS) A4 (210X297 mm) (Please read the back (Please fill in this page again)

Printed by the Central Bureau of Associate of the Ministry of Economic Affairs and the Consumer Cooperative 593305 A7 B7 V. Description of the invention (42) Botrvosphaeria obtusa IMI 03 8560, Caloneotria decora ATCC 14767, Chaetomium cochliodes ATCC 10195 Corynespora cassiicola ATCC 16718, Cunninghamella blakesleeana ATCC 8688a, Cunninghamella echinulate ATCC 3655, Cunninghamella elegans ATCC 9245, Curvularia clavata ATCC 22921, Curvularia lunata ACTT 12071, Cylindrocarpon radicicola ATCC 1011, Epicoccum humicola ) ATCC 12722, Gongronella butleri, ATCC 22822, Hvpomyces chrvsospermus, Mor_tierella isabellina ATCC 42613, Mucor mucedo ATCC 4065, gray blue and white Mold (Mucor griseo_cyanus) ATCC 1207A, Myrothecium verrucaria ATCC 909 5. Coral soil mycelia (Nocardia corallina), Kanas expansion: Green machine: (P aecilomvces carneus) ATCC 46579, Penicillum patulum (ATCC 24550), Pithomvces atro- olivaceus, IFO 6651, Pithomyces cvnodontis ATCC 26150, Pycnosporium sp · ATCC 12231, Saccharopolyspora erythrae, ATCC 1 1635, Seputeonium chrysospermum, ATCC 13 2. Two-color Stachvlidium bicolor ATCC 12672, Streptomyces hygroscopicus ATCC 2743 8. Streptomyces hygroscopicus-45- This paper is in accordance with China National Standard (CNS) A4 (21 〇X297) Mm) (Please read the notes on the back before filling this page) Order 593305 A7 B7 _ V. Description of the Invention (43) (Streptomyces purpurascens) ATCC 25489, Syncephalastrum racemosum ATCC 18192, Pear-shaped Satsuma Thamnostylum piriforme ATCC 8992, Thielavia terricola, ATCC 13 807, and Vertici llium theobromae) ATCC 12474 o Printed by the Central Bureau of Commerce of the Ministry of Economic Affairs and the Consumer Cooperative (please read the notes on the back before filling out this page) Additional organisms that can be expected to show activity on 11 π-hydroxylation include jg_ 菲迪 可Cephalosporium aphidicola (Phytochemistry (1996), 42 (2), 411-415), Cochliobolus lunatas KJ. Biotechnol. (1995), 42 (2), 145--150) , Tieghemella orchidis, (Khim.-Farm. Zh. 9 (186), 20 (7), 871-876), Tieghemella hvalospora ) (Khim. Farm. Zh. (1986), 20 (7), 871-876), Monosporium olivaceum (Acta Microbiol. Pol., Ser. B. (1973), 5 ( 2), 103-110), Aspergillus ustus, (Acta Microbiol · Pol ·, Ser_B. (1973), 5 (2), 103-110), Fu_sarium graminearum (Acta Microbiol · Pol, · Ser. B. (1973), 5 (2), 103-110), Verticillium glaucum (Acta Microbiol. Pol., Ser. B. (1973), 5 (2 ), 103-110), and Rhizopusnigricans (J. Steroid Biochem. (1987), 28 (2), 197-201) 〇 For the hydroxylation of Conreno or other substances of formula XIII, preparation of production-scale fermentation, including A seed fermentor, or a cell fermentation system of one or two or more seed fermenters, prepares an inoculum of cells. Introduce the working material spore suspension together with the nutrient solution for cell growth into the first -46- This paper size applies Chinese National Standard (CNS) A4 specification (210X297 mm) 593305

、 Explanation of the invention (44 Employees' cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs print a variety of seeds into the fermenter. If the production system wants or needs; more than the volume produced in the first seed fermenter, you can borrow ":: T species: the progress of the remaining fermenters in the Γ series, gradually increasing the volume of the inoculum with the y formula. It is best to inoculate the seed fermentation system, which is sufficient to produce fermentation, The reaction started quickly and reached a relatively short production batch cycle, which was dependent on the active product and viable cells of the A ^ shield and the production fermenter. Sowing on the number of containers in a series of seed fermenters How much, the second and subsequent seed fermenters are preferably of a certain size, so that the degree of dilution in each step of the row is substantially the same. In each seed fermenter, the initial inoculum The dilution is about the same as that in the production fermenter. Conrano or other substances of formula XIII are charged into the production fermenter together with the inoculum and nutrient solution, and the hydroxylation reaction is operated from there.The spore suspension introduced into the seed fermentation system is a small raw tile of working material spore suspension obtained from most vials constituting the working material cell bank, which is stored under low temperature conditions before use. The working material cell bank is below The method is listed below, successively derived from the main raw material cell bank. Obtain from an appropriate source initially suspended in liquid culture medium, such as, for example, a physiological saline solution, a nutrient solution, or a surfactant solution (eg, a nonionic surfactant, such as about 0.001 20% by weight of spore samples in Tween 20), such as ATCC, and distribute the suspension in culture plates, each plate containing a solid nutrient mixture, usually a non-digestible polysaccharide such as agar Based on this, the spores are allowed to multiply there. The solid nutrient mixture preferably contains between about 0.5% and about 5% by weight of glucose, and the Chinese National Standard (CNS) A4 specification (210X297) is applied at about -47-degree. Mm) (Please read the notes on the back before filling in this page)

593305 A7 B7 V. Description of the invention (45 Employees' cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs of the People's Republic of China printed a nitrogen source, such as a gland, between about 0.05% and about 0.5% by weight. % Of phosphorus sources, such as ammonium phosphate or alkali metal phosphate, such as dipotassium hydrogen phosphate, yeast lysates or extracts (or other amines) between about 025% and about 2.5% by weight Source of acid, such as meat extract or brain heart infusion), agar or other non-digestible polysaccharides between about 1% and about 2% by weight. A solid nutrient mixture may be included as needed, and / or contained in about 〇 · 丨% by weight to about 5 wt./malt extract between / 〇. The pH of the solid nutrient mixture is preferably between about 5.0 · about 7 · 0, by alkali metal as needed It can be adjusted by hydroxide or orthophosphoric acid. The solid growth medium that can be used is as follows: 1 · Sol® Medium # 1 ·· 1% glucose, 0.25% yeast extract, 0.3% K2HP04 and 2% agar (Bacto); Adjust pH to 6.5% with 20% NaOH 2% (Bacto), 1% yeast extract (Bacto), 2% glucose and 2% agar (Bacto); adjust pH to 5 with 10% H3PO4. 0.1% gland (Bacto), 2% malt extract (Bacto ), 2% glucose and 2% agar (Bacto); pH 値 adjusted to 5. 3.5% brown sugar paste, 0.5% corn pulp, 0.25% glucose, 0.25% NaCl and 0.5% KH2P 〇4, pH 値 adjusted to 5.8. Difco agar agar (low pH 値). For the future needs of the main raw materials, you can choose to expand the main raw material cells 2 · Solid medium # 2 • solid sad medium # 3: 4 · Liquid medium 48- This paper size is in accordance with Chinese National Standards (CNs) A4 (210X 297 mm) (Please read the precautions on the back before filling this page) 593305 Printed by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7 B7 5 Description of the invention (46) The number of agar culture plates used in the bank, but usually about 15 to about 30 culture plates are prepared. After a suitable growth period, for example, to 10 days, in the presence of an aqueous vehicle Scrap the culture dish, usually saline or buffer solution, to harvest the spores The obtained main raw material suspension is divided into several vials, for example, 1 ml each in a plurality of 15 liter vials. To prepare a spore suspension of working raw materials used in research or production fermentation operations, The contents of one or more of these second-generation vials of the main raw materials were distributed on an agar culture plate and cultured in the manner described above for preparing the spore suspension of the main raw materials. When routine manufacturing operations are completed, as many as 100 to 400 plates can be used to produce second-generation working materials. Scrape each culture dish into different working material vials, each vial usually containing 1 ml of the inoculum produced. For permanent preservation, the main raw material suspension and the second-generation inoculum can be advantageously stored in the vapour area of a cryogenic storage container containing a liquid or other cryogenic liquid. In the method illustrated in Fig. 1, a liquid growth medium is prepared, which includes a nitrogen source such as a gland, a yeast derivative or equivalent, glucose, and a phosphorus source such as a phosphate. In this medium in a seed fermentation system, spores of microorganisms are cultured. The preferred microorganism is ^ nrrl 4 () 5 (ATCC 1 8500). The seed material thus produced is then introduced into a production fermenter together with the enzyme of formula XI11. The fermentation broth is agitated and aerated for a time sufficient to allow the reaction to proceed to the desired degree of completion. The culture medium of the seed fermenter's is made up of a liquid mixture, which is inherently there. Glucose between about 0.5% to about 5% by weight, nitrogen between about 0.05% to about 5% by weight Sources, such as glands, are approximately -49- This paper size applies Chinese National Standard (CNS) A4 specifications (2 丨 OXM7 mm) (Please read the precautions on the back before filling this page), 11 II--I- I -. 593305 Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs A7 B7 V. Description of the invention (47) 0 · 0 5 wt% to about 0.5 wt% phosphorus sources, such as ammonium phosphate or alkali metal phosphate, Like a monovalent ammonium phosphate or dipotassium hydrogen phosphate, a yeast lysate or extract (or other amino acid source, such as a wine tank) between about 0.25% to about 2.5% by weight. Agar or other non-digestible polysaccharides between about 1% and about 2% by weight. Particularly preferred seed growth media contain nitrogen sources between about 0.05% to about 5% by weight, such as glands, autolysed yeasts or yeasts between about 0-25% by weight and about 2.5% by weight The extract 'glucose' between about 0.05 to about 5% by weight and a source of phosphorus between about 0.05 to about 0.5% by weight, like ammonium monovalent phosphate. By using other preferred seed media, a particularly economical method of operation is obtained, which comprises between about 0.5 and about 5% by weight corn syrup, and between about 0.5 and about 2.5% by weight Between autolyzed yeast or yeast extract, glucose between about 0.5% and about 5% by weight, and between about 0.5% by weight. / () To about 0.5% by weight of monovalent ammonium phosphate. Corn pulp is a particularly economical source of proteins, peptides, carbohydrates, organic acids, vitamins, metal ions, trace substances and phosphates. Instead of or in addition to corn syrup, syrup derived from other grains can also be used. The pH of the medium is preferably adjusted to a range of about 5-0 to about 7.0, for example, by adding an alkali metal hydroxide or orthophosphoric acid. Where corn slurry is provided as a source of nitrogen and carbon, it is best to trim maggots to a range of about 6.2 to about 6.8. For a medium including glands and glucose, it is preferable to adjust the pH to between about 5.4 and about 6.2. The growth media that can be used in seed fermentation include: 1. Medium # 1: 2% gland, 2% autolyzed yeast (or yeast extract), and 2% grapes (read the precautions on the back before filling this page)

-50- Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs 593305 A7 ------ _ B7 V. Description of the invention (48) Sugar; adjust pH 値 to 5.8 with 20% NaOH. 2 · 杧 养 基 # 2: 3% corn slurry, 1.5% yeast extract, 0.3% ammonium monovalent gallate and 3% glucose; adjust pH to 6.5 ° with 20% NaOH will be obtained from the usual Microbial spores in vials containing approximately 109 spores per liter of suspension were introduced into the medium. Optimal productivity for seed reproduction is obtained when dilution with growth medium at the beginning of seed culture does not reduce the density of the spore population to below 107 per milliliter. Preferably, the spores are cultured in a seed fermentation system until the volume of filled mycelia (PMV) is at least about 20%, and more preferably from 350/45 to 45%. Because the circulation in a seed fermentation container (or any number of containers that make up the seed fermentation rank) depends on the initial concentration in the container, you may want to prepare one or two seed fermentation stages to speed up the entire process. However, it is best to avoid the use of more than two seed fermenters in a row, because if the seed fermentation is done through an excessive amount of stages, the activity may be compromised. Seed culture fermentation is performed in a temperature range of about 2 3 to about 37 under agitated conditions, preferably in a range of about 24 ° C to about 28χ :. The culture obtained from the seed fermentation system is introduced into the production fermenter together with the production growth medium. In a specific embodiment of the present invention, Conreno sowed or other SXIIIi enzymes are used as the enzymes of the reaction. Preferably, the enzyme-receiving substance is added to the production fermenter in a form of 0% to 30% by weight in a growth medium. To increase the surface area available for the hydroxylation reaction, by introducing the enzymatic substance of formula χπι into the fermentation -51- This paper size applies the Chinese National Standard (CNS) ^ 4 specifications (2ι〇 &gt; &lt; 297) Policy) (Read the precautions on the back before filling in this page)

Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs 593305 ______ B7 V. Description of the invention (49) Before passing the device through an external grinder to reduce the size of the enzymatic radon. Sterile nutritional feed ingredients containing glucose were also introduced, and a second type of sterile nutrition containing yeast derivatives, such as autolyzed yeast (or equivalent amino acid formula, based on another source, such as a wine tank) Solution. The culture medium consists of an aqueous mixture containing the following ingredients: glucose between about 0.5% to about 5% by weight, nitrogen sources between about 0.05% to about 5% by weight, such as glands, at Sources of phosphorus between about 0.05% to about 0.5% by weight, such as acetic acid or metalloate, such as hydrogen diacetate, at about 0.25% to about 2% · Yeast lysates or extracts (or other amino acid sources such as wine tanks) between 5% by weight, agar or other non-digestible polysaccharides between about 1% and about 2% by weight. Extraordinarily good production growth medium containing a nitrogen source between about 0.05% by weight and about 5% by weight, such as a gland, between about 0.25% by weight and about 2.5% by weight Autolyzed yeast or yeast extract, glucose between about 0.5% to about 5% by weight, and a source of phosphorus between about 0.05% to about 0.5% by weight, like one Valent ammonium phosphate. Other preferred production media contain corn syrup between about 0.5% and about 5% by weight, autolyzed yeast or yeast extract between about 0.25% and about 2.5% by weight, and about 0.5 to about 5% by weight ammonium glucose, and monovalent ammonium phosphate between about 0.5 to about 0.5% by weight. It is preferable to adjust the pH of the fermentation medium to be adjusted by the above-mentioned methods for the seed fermentation medium having the same preferable pp range as the gland / glucose-based medium and the corn-based medium, respectively. Available biotransformation growth media are listed below: • 52- This paper size applies to China National Standard (CNS) A4 (210X297 mm) &quot; (Please read the precautions on the back before filling this page)

593305 Printed by the Consumer Standards Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs A7 B7 V. Description of the Invention (50) 1. Medium # 1: 2% gland, 2% autolyzed yeast (or yeast extract), and 2% glucose; 20% NaOH adjusted the pH to 5.8. 2 • Medium # 2: 1% gland, 1% autolyzed yeast (or yeast extract) and 2% glucose; adjust pH to 5.8 with 20% NaOH. 3 Medium # 3: 0.5% gland, 0.5% autolyzed yeast (or yeast extract) and 0.5% glucose; adjust pH to 5.8 with 20% NaOH. 4 · Medium # 4: 3% corn slurry, 1.5% yeast extract, 0.3% ammonium phosphate and 3% glucose; pH% was adjusted to 6.5 with 20% NaOH. 5 · Medium # 5: 2 · 55% corn slurry, 1 · 2 75% yeast extract, 0 · 2 55% monovalent ammonium phosphate and 3% glucose; pH pH was adjusted to 6.5 with 20% NaOH. 6 · Medium # 6: 2.1% corn slurry, 1.05% yeast extract, 0.21% ammonium phosphate and 3% glucose; pH 値 was adjusted to 6.5 with 20% NaOH. Non-sterile Conley and sterile nutrient solutions are supplied to the production fermentation in a five-to-20, preferably ten to fifteen, and preferably approximately equal ratio in a production batch cycle Device. Advantageously, prior to inoculation with the seed fermentation broth, initially a concentration sufficient to establish between about 0.1% to about 3% by weight, preferably about 0.5% to about 2% by weight The content of the concentration is introduced into the substrate, and then added periodically, conveniently every 8 to 24 hours, to a cumulative ratio between about 1% by weight and about 8% by weight. Where additional enzymes are added every 8 hours, the total amount added is slightly lower than the case of adding enzymes only on a daily basis, such as -53- This paper size applies Chinese National Standard (CNS) A4 Specifications (210X297 mm) (Please read the notes on the back before filling this page)

593305 Stamp printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs Α7 Β7 V. Description of the invention (51) 0 · 25 5 to 2 · 5 wt%. In the latter case, the cumulative Conranow addition amount that may be required is in the range of 2% to about 8% by weight. The supplemental nutritional mixture supplied during the fermentation reaction is preferably a concentrate, for example, containing sterile glucose between about 40% to about 60% by weight, and between about 16% to about 32% by weight A mixture of sterile yeast extracts or yeast derivatives (or other amino acid sources) of other sterile origin. Because the enzymes supplied to the fermenter produced in Figure 1 are non-sterile, antibiotics are added to the fermentation broth to control the growth of undesired organisms. Antibiotics such as cefalexin) have no adverse effects on growth and biotransformation. It is preferred to 'introduce these antibiotics into the fermentation broth at a concentration between about 0.004 wt% to about 0.02 wt% based on the total content of the broth', including, for example, re- Based on the total broth content, between about 20000% by weight f% to about 0. 006% by weight of Conomycin sulfate, between about 0.002% by weight and about 0.00%. Tetracycline H c ^ between 6% by weight, and / or cephalosporin between about 0.001% by weight and about 3,000% by weight. Usually, the batch fermentation cycle is about 80-60 hours. Therefore, one serving of the enzyme enzyme and the nutrient solution of formula X is generally added every 2 to 10 hours, preferably every 4 to 6 hours. Advantageously, the antifoaming agent is also incorporated into the seed fermentation system and into the production fermenter. In the method of FIG. 1, the inoculum filled in the production fermenter is based on the volume of the total mixture in the fermenter, preferably about 0.5 to about 7/0, and more preferably about 1 To about 2%, and the glucose concentration is preferably maintained between about 0.01% by weight and about 10% by weight, preferably (please read the precautions on the back before filling in this page)

-54- 593305 Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs A7 Β7 V. Description of the Invention (52) About 0 · 0 2 5 wt% to about 0 · 5 wt%, more preferably about 0 · 0 Between 5% by weight and about 0.25% by weight, and for periodic addition, it is preferably based on the total loading in batches, with a portion of between about 0.05% by weight and about 0.25% by weight. It is convenient to control the fermentation temperature in the range of about 20 t to about 37 ° C, preferably about 24. (: To about 28m, but lowering the temperature during the reaction period may also be desirable, for example, to increase by 2 ° F to maintain the filled mycelium volume (PMV) below about 60%, preferably below about 50% 'and thereby avoid the viscosity of the fermentation broth, which prevents satisfactory mixing. If the biocluster grows and extends above the liquid surface, the enzymatic substance retained in the biocluster can complete the reaction zone, And become unavailable for the hydroxylation reaction. For productivity, it is desirable to reach 30 to 50% PMV within the first 24 hours of the fermentation reaction, preferably 35% to 45%, but it is best to try to make Subsequent conditions control further growth within the above-mentioned limits. During the reaction, the pH of the fermentation medium is controlled to be between about 5.0 to about 6.5, preferably between about 5.2 to about 5.8, And agitating the fermenter at a speed between about 400 to about 800 rpm. By inflating in batches at about ο "to about 1.0 vv m, a dissolved oxygen amount of at least about 10% saturation is achieved, And maintain the pressure in the headspace of the fermenter at about atmospheric pressure Between about 1.0 bar, preferably about 0.7 bar. The stirring rate can also be increased as needed to maintain a minimum dissolved oxygen amount. Advantageously, the dissolved oxygen amount is maintained above 10%, in fact As high as 50%, it can promote the conversion of the enzyme. Maintaining p Η 値 within the range of 5.5 ± 0.2 is also most suitable for biotransformation. By adding general antifoam as needed To control the foaming. After adding all the enzymes, it is best to continue the reaction until the product of formula VIII on the residue -55- This paper size applies the Chinese National Standard (CNS) A4 size (210X 297 mm) (please read the back first) Note before filling out this page}

593305 The A7 B7 printed by the Consumer Cooperative of the Central Standards Bureau of the Five Ministry of Economic Affairs, the invention description (more than 53 unreacted formula XI11, the molar ratio of the enzyme is at least about 9 to 1. Such a change can be between 80 and 16 〇The above specified batch cycle was achieved. It has been found that the high conversion related to the initial nutrient consumption lower than the initial filling amount, by controlling the aeration rate and agitation rate, to avoid splashing liquid broth by enzymes In the method of Figure 丨, the nutrient content is consumed and then maintained at an initial filling amount of not more than about 60%, preferably about 50%; and in the method of Figures 2 and 3, the nutrient content Reduced to and maintained at no more than about 80% of the initial fill, preferably about 70%. The inflation rate is preferably no more than 1 vvm, more preferably in the range of about 0-5 vvm; and The agitation rate is preferably not more than 600 rprn. A particularly preferred method for preparing a compound of formula VIII is illustrated in Figure 2. The preferred microorganism is still 405 (ATCC 18500). In this method, the growth medium is preferably included in About 0.5% by weight Corn syrup between about 5 wt.%, Glucose between about 0.5 wt.% And about 5 wt.%, Yeast extract between about 0.1 wt.% And about 3 wt.%, And about 0.05% to about 0.5% by weight of ammonium phosphate. However, other growth-producing media described herein can also be used. Seed cultures are basically prepared in the manner described in Figure 丨 and used herein Any seed fermentation medium described in. In a mixer, prepare a suspension of non-micronized Conorno or other enzymes of the formula X in the growth medium in a sterile manner, preferably at about 0% by weight. To a relatively high concentration of about 30% by weight of the enzyme. Preferably, the sterile product may include sterilizing the suspension after mixing or using pasteurization. Paper size applies Chinese National Standard (CNS) A4 specification (2I0X297 mm) (Please read the precautions on the back before filling this page}

593305 A7 B7 V. Description of the invention (54) The total amount of aseptic enzyme-containing suspension required for the batch, which will be introduced into the production system through regular chain supply. The size of the particles is cut and held on the spring, so avoiding the "wet grinding" reduces the need for a pasteurizer outside the pasteurizer instead of sterilizing. σ & ^ Pasteurella method to achieve aseptic conditions, coagulation ^^, I want to use the male cutting pump, a positive control group of kU, size. Basically, in a similar manner as described above, the bacterial growth-promoting oxygen and glucose solution are introduced into the production fermentation cry. All products supplied to the production fermenter are sterilized before introduction, so antibiotics are not required. It is better to print from the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. In the method 0 of Figure 2, the inoculum is introduced into the production fermenter at a ratio of about 5% to about 7%. The temperature is between about 20 C and about 37 ° C, preferably between about 24 T: and about 28 C, and by, for example, introducing ammonia, an aqueous ammonium hydroxide solution, an aqueous alkali metal hydroxide solution, or orthophosphoric acid The pH is controlled to be between about 4.4 and about 5.5, and preferably between about 5.3 and about 5.5. As in the method of Figure i, it is best to adjust the temperature to control the growth of the bioclum so that the PMV does not exceed 5 5 -60%. The initial filling of glucose is preferably at about! It is preferably between about 2.5% and about 4% by weight, preferably between about 2.5% and 3.5% by weight, but it is also allowed to gradually fall below about 1.0% by weight during fermentation. 1 重量 ％ 至 To a regular supply of supplemental glucose between about 0.2% by weight / g to about 10% by weight, depending on the batch loading, in order to maintain the glucose concentration in the fermentation zone at about 0.1% by weight to Within the range of about 1.5% by weight, more preferably between about 0.25% by weight and about 0.5% by weight. May 57- This paper size applies to Chinese National Standards (CNS) A4 (210X 297 mm) 593305 A7 B7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (55 Supplement nitrogen and phosphorus sources together with glucose as needed However, because the total filling capacity of Connor is determined at the beginning of the batch cycle, it is also possible to introduce nutrient-containing nitrogen and tritium necessities at this time, allowing only glucose solution to be used to supplement it during the reaction. The rate and nature of the change are clearly variable. Moderately agitated to promote the transfer of the mass between the solid substrate and the liquid phase. However, a low-shear beater should be used to prevent the microbial bone marrow phosphorous. Degradation. The optimum agitation speed varies within the range of 200 to 800 rpm, depending on the viscosity of the broth, the oxygen concentration, and the mixing conditions affected by the container, baffle, and agitation mechanism. Usually a better agitation The speed is in the range of 350-600 rpm. It is best to stir the stirrer to provide the axial down extraction function in order to assist the good mixing of the fermented biological mass. Each batch It is good to inflate at a rate of about 0.3 to about 1.0 vvm, more preferably 0.4 to 0.8 vvm, and the pressure of the space on the liquid surface of the fermenter is preferably about 0.5 to about 0.5 Between 1.0 bar. Temperature, agitation, aeration and back pressure are preferably controlled in order to maintain the amount of dissolved oxygen in the range of at least about 10% by volume during the biotransformation. The total batch cycle is usually around 100 To about 40 hours. Although the principle of operation of the method of Figure 2 is based on the introduction of substantially the entire Conranow loading at an early stage, it will be clear that the growth of fermented broth may be loaded in most Conranos Completed previously. A portion of Conranox can also be added later in the batch if necessary. However, at least approximately 75% of the sterile Conranault loading should normally be introduced into the conversion fermentor within 48 hours of starting fermentation In addition, it is also hoped that at the beginning of fermentation, or at least within the first 24 hours, at least about 25% by weight of Conreno will be 'introduced' in order to promote biotransformation enzymes. 210X297 mm) (Please read the precautions on the back before filling out this page) — · Γ 593305 A7 B7 V. Description of the invention (56) The production of primes (groups). In the better method shown in Figure 3, Before introducing the inoculum, the entire batch of filling and nutrient solution is sterilized in the production fermentation container. The nutrient solution that can be used, and the better among them, are basically the same as those in the method of Figure 2. In a specific embodiment of the invention, the shearing action of the blender is to destroy the enzymatic agglomerates that are liable to form during sterilization. It has been found that if the average particle size of Conorno is less than about 200 microns and at least 75% by weight of the particles Below 240 micrometers, the reaction can proceed satisfactorily. It has been found that the use of a suitable blender, such as a disc-shaped steam turbine beater, at a suitable speed in the range of 2000 to 800 rpm, using a peripheral speed of at least about 400 cm / s, provides sufficient to maintain such particle size. The characteristic shear rate is prone to agglomeration regardless of the sterilization effect in the production fermenter. The remaining operations of the method of FIG. 3 are basically the same as those of the method of FIG. 2. The methods of Figures 2 and 3 provide several different advantages over the method of Figures 丨. A special advantage is the willingness to use low-cost nutrient substrates, such as corn slurry. However, it is clear that more advantages are the reduction of the need for antibiotics, the simplification of the supply process, and the batch sterilization of enzymes of Conorno or other type XIII. Other special advantages are the ability to use simple glucose solutions without having to replenish complex nutrient solutions during the reaction cycle. In the methods described in Figures 1 to 3, the product of Figure viii is a crystalline solid that can be separated from the reaction broth together with the biological mass by filtration or low-speed centrifugation. Alternatively, organic solvents can be used to extract products from the entire reaction broth. The product of formula v 111 is recovered by solvent extraction. In order to maximize the recovery, the extraction solvent is used to treat the liquid filtrate and the biological mass. -59 · This paper size is applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) (Please read the precautions on the back before filling this page) Ding printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 593305 printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7 —- —___ B7 V. Description of the invention (57) ~ Filter paper or Centrifuge the cake, but usually -95% of the product stays with the biomass. Extraction is usually carried out using hydrocarbons, esters, vaporized hydrocarbons and ketone solvents. The preferred solvent is ethyl acetate. Other representative suitable solvents include toluene and methyl isopropyl ketone. For extraction from the liquid phase, it is convenient to use a solvent in a volume approximately equal to the volume of the reaction solution it encounters. To recover the product from the biomass, float the biomass 54; the aphrodisiac is taken in a large excess of solvent relative to the initial enzymatic loading, such as 5 grams per gram of initial Conorno 0 to 100 ml of &lt; solvent, and preferably the resulting suspension is refluxed for a period of 20 minutes to several hours, to ensure that the product moves from the depth of the biomass and the pores to the solvent phase. Subsequently, the biological mass is removed by filtration or centrifugation, and preferably the filter cake is rinsed with both fresh drops and deionized water. The aqueous solution and solvent are then mixed and the phases are allowed to separate. The product of formula VIII is recovered by forming crystals from this solution. For maximum yield, contact the mycelium with fresh solvent twice. After precipitation, the liquid phase was allowed to separate completely and the product was recovered from the solvent phase. It is best to remove the solvent under vacuum until crystallization begins, and then the concentrated extract is cooled to 0OS2 (rc, preferably 101 to 15 c 'for a period of time sufficient to begin crystallization and growth, Usually 8 to 12 hours. The method of Figure 2 and especially the method of Figure 3 are particularly good. These methods operate at low speed and bear the responsibility of tightly controlling the parameters such as temperature, oxygen and oxygen. In addition It is easy to maintain sterile conditions without relying on antibiotics. The biotransformation process is exothermic, so that a jacketed fermenter or this paper should be used. (Please read the precautions on the back before filling out this page)

1T 593305 A7 B7 Consumer Cooperative Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs. 5. Description of the Invention (58) is a cooling coil in the product fermenter to remove heat. Alternatively, the reaction broth may be circulated through an external heat exchanger. By measuring the oxygen potential of the reaction in the broth, the rate of introducing air into the reactor is adjusted so that the amount of dissolved oxygen is maintained at a level of at least about 5 vol%, preferably at least about 0 vol%, which is sufficient to provide The energy required for the reaction, and to ensure that glucose is converted to (: 0 2 and H 2 0. It is best to control the pH to be between about 4.5 and about 6.5. In each of the different enzymes of formula XIII, the 11-light group In the chemical reaction method, the productivity is limited by the mass transfer from the solid substrate to the liquid phase, or the phase interface of the reaction occurring at a distance. As shown above, as long as the average particle size of the substrate Reduced below about 300 microns, and at least 75% by weight of the particles are less than 24 microns, productivity is not significantly limited by the mass transfer rate. However, these may be further promoted in certain embodiments. The productivity of the method, which provides the fact that Conreno or other enzymes of formula XIII are loaded into a production fermenter in an organic solvent. According to one option, the enzymes are dissolved in an immiscible water In solvent It is mixed with an aqueous growth medium inoculum and a surfactant. Immiscible solvents that can be used include, for example, DmF, DMSO, (: 6-(: 12 fatty acids, n-alkanes, vegetable oils, sorbitan, and Aqueous surfactant solution. The filling-emulsion reaction system is instigated, with an extended interface area # for T-enzyme to transfer the mass from the organic liquid phase to the reaction site. The first choice &amp; @ 始 将 受 酶 质Dissolved in water-miscible solvents' like acetone, acetone, ethyl acetate, or glycerin, which has a known concentration in water. ^ ^ ^ Preparation of the first enzyme at the temperature: female ,, and ^ ^ t ^ combined shoes, increase solubility, thereby increasing the size of the paper on the solution. Read the notes on the back and fill in this page) Order -jim -61- Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 593305 A7 ______________B7 V. Description of the invention (59) ~ The amount of the enzyme into the reactor in the form, And finally promote the load on the reactor. The enzymatic solution is loaded into the production fermentation reactor together with the relatively cold aqueous filling material, which includes the growth medium and the inoculum.殿. However, 'under the conditions of generally supersaturated and moderately active agitation, the nucleus opens; the formation effect is better than crystal growth' and forms very fine particles with high surface area. Such high surface area promotes the clumps in the liquid Transfer between the phase and the solid substrate. In addition, the presence of a water-miscible solvent also promotes the equilibrium concentration of the substrate in the aqueous liquid phase. As a result, productivity is increased. Although microorganisms may not be tolerant in the liquid phase High concentration solvents, but for example, ethanol concentrations ranging from about 3% to about 5% by weight can be effectively used. The second option is to dissolve the enzyme in the aqueous cyclodextrin solution. Cyclodextrins which may be exemplified include hydroxypropyl-fluorene · cyclodextrin and methyl-cyclodextrin. The molar ratio of the enzymatic substance to be cyclodextrin can be about 丨: 丨 to about 丨: 丨 · 5. The enzymatic substance to be digested: cyclodextrin. The enzymatic: cyclodextrin mixture can then be added to the biotransformation reactor in a sterile manner. 1 1 per 每 Conrad and other 1 1 α _ the products of the abasification (formula V jjg and VIIIA) are novel compounds, which can be filtered by the reaction medium, and from the organisms collected on the filter medium The product is separated from the pellet by extraction using conventional organic solvents such as ethyl acetate, acetone, toluene, vaporized hydrocarbons, and methyl isobutyl ketone. The product of formula VIII can then be recrystallized from an organic solvent of the same type. The compound of formula νπι has as a preparation formula; [compounds, especially intermediates of eight compounds of formula-62- This paper size applies to China (CNS) A4 specifications (210xΥ97 mm) --- (Please read the note on the back first Please fill in this page again. Bookmark 593305 A7 -_______ B7 V. Actual price of invention description (6〇) A better compound of formula VIII, in which -AA- and · Β-Β_ is -CH2- CHy, R3 is Hydrogen, low-carbon alkyl or low-carbon alkoxy, and r8 and r9 together form a 20-spiroxane ring:

A compound of formula VIIIA. It is best to react the compound of formula V111 with a cyanide ion source under alkaline conditions to produce an enamine compound of formula VII according to the method of plan 1. (Please read the precautions on the back before filling this page)

R3 .β

Order

VII here where-A-A-, R3, -B-B-, R8 and R9 are as defined above. The enzyme is equivalent to formula VIIIA, the product of formula VIIA. Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs.

63- VII a This paper size applies the Chinese National Standard (CNS) A4 specification (210X 297 mm) 593305 A7 B7 V. Description of the invention (61 where -A-A-, _ B _ B are defined.

R Y1, γ2, and X are the same as in Formula XIII. The 11α-hydroxy group of Formula VIII printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs is subjected to enzymatic cyanation, which can be sourced with cyanide ions, such as The cyanohydrin, most preferably acetone nitrogen cyanide, is completed by reaction in the presence of a base and an alkali metal salt, and most preferably Li (R). In addition, cyanation can also be accomplished by using an alkali metal cyanide in the presence of an acid without the need for a cyanohydrin. In the ketocyanohydrin method, the reaction is run in solution, preferably using an aprotic polar agent such as dimethylformamide or dimethylsulfoxide. Forming amidine requires at least two sources of cyanide ion per mole of enzymatic substance, and it is best to use a slight excess of the source of cyanide. The base is preferably a nitrogen-containing base such as dialkyl, trisamine, alkanolamine, pyridine or the like. However, it is also possible to use inorganic bases, such as alkali metal carbonates or alkali metal hydroxides. Preferably, the enzymatic substance of formula VII] [is present at a ratio of about 20% to about 50% by weight from the beginning, and the base is between 0.5 and two equivalents per equivalent of the enzyme. The proportion exists. The reaction temperature is not strictly limited, but productivity can be promoted by operating at an elevated temperature. Therefore, for example, using triethylamine as a test, the reaction is advantageously operated in a range of about 80 to about 90. At such temperatures, the reaction proceeds to completion in about 5 to 20 hours. When diisopropylethylamine is used as the base and the reaction is run at 105 'the reaction is completed at 8 hours. During the end of the reaction, remove the solvent under the water, dissolve the remaining oil in water, and dilute the hollow with dilute acid (please read the precautions on the back before filling this page). Applicable to Chinese National Standard (CNS) A4 specification (21 × 297 mm) 593305 A7-· ~ -_____ Β7 V. Description of invention (62) Printed to P Η 7 by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs, better It is hydrochloric acid. The product was precipitated from the solution, and then A was washed with steam and air-dried. The released He / can be stripped with inert gas and stopped in alkaline solution. The dehydrated precipitate is dissolved in aerobic or other suitable solvents, and then extracted with a concentrated acid such as 6N HC1. The extract is neutralized to 7 by adding an inorganic base, preferably a metal hydroxide, and cooled to a temperature range of generation. The resulting precipitate is washed and dehydrated and then recrystallized from a suitable solvent, such as acetone, to produce a product of formula νπ suitable for use in the next step of the method. Alternatively, the reaction can be operated in an aqueous solvent system including a water-miscible organic solvent such as methanol, or in a two-phase system including water and an organic solvent such as ethyl acetate. In this option, the reaction solution can be diluted by water, followed by extraction of the product with an organic solvent such as dichloromethane or chloroform, and then the use of a concentrated inorganic acid, such as 2 reverse hci, from the organic extract Back extraction to recover product, see US Patent No. 3,200,113. Depending on the choice, the reaction can also be run in a water-miscible solvent such as dimethylformamide, dimethylacetamide, N-methylpyrrolidone, or dimethylsulfoxide to After diluting the reaction product solution with water and making it qualitative, for example by adding a metal test salt, it is then cooled to zero. . To 10 t, which caused the product to precipitate. It is best to stop the system with an alkali metal hypochlorite or other agent that is effective in preventing cyanide evolution. After filtration, washing with water, the precipitated product is suitable for use in the next step of the method. According to more choices, the formula can be used in the presence of a proton source (please read the precautions on the back before (Fill in this page) Yi'-65- This paper size is applicable to Chinese National Standard (CNS) A4 specifications (210X297, 593305, printed by the Consumers' Cooperatives of the Central Standards Bureau, Ministry of Economic Affairs, printed by A7 __ B7 V. Description of invention (63) VIII enzyme React with an excess of an alkali metal cyanide, preferably NaCN, in an aqueous solvent including an aprotic, water-miscible polar solvent such as dimethylformamide or dimethylacetamide To produce the amidine product of formula V 丨 J. The source of the proton is preferably an inorganic acid or a complex acid of ci to C5, particularly sulfuric acid. The exception is when the reagent for cyanation is in DMF For commercial LiCN, it is not necessary to add a separate source of protons. Cyanide ions are preferably charged into the reactor at a ratio of about 2.05 to about 5 mole equivalents per equivalent of enzymatic substance. Acids or other proton sources, Helps H CN to cross the 4, 5 and 6, 7 double bond addition, and preferably exists in a ratio of at least one mole equivalent per enzyme equivalent; however, the reaction system should be maintained by exceeding The excess of acid is tested for metal cyanide 'to maintain it as a test. The reaction is preferably completed at a temperature of at least about 7 5 ° c, usually 60 ° c to 100 ° C, for about! To about 8 hours' It is preferably about 1.5 to about 3 hours. At the end of the reaction, the reaction mixture is cooled, preferably to about room temperature; and by acidifying the reaction mixture and mixing with cold water, it is preferably about ice At the temperature of the bath, the product amidine was precipitated. Salty believed that acidification turned off 17 _ lactone, which was easily opened under alkaline conditions where cyanidation prevailed. The same acid was used as it appeared during the reaction It is convenient to acidify the reaction mixture, preferably sulfuric acid. It is best to add water in a ratio between about 10 to about 50 mole equivalents per mole of product. The compound of formula VII is a novel compound and has As a compound of formula In particular, the actual valence of the intermediate of the compound of the formula IA. Compared with the compound, it is in line with · Α 一一 H2_CH2 _ "= ^^ -66- This paper size applies the Chinese National Standard (CNS) A4 specification (21 〇χ 297 mm ) (Please read the notes on the back before filling this page)

, 1T 593305 A7 B7

Description of the invention (64, low-carbon alkyl or low-carbon alkoxy, and R8 and R9 together form a 20-spiroxane ring:. XXXI industrial foot compound IIA. The best compound ^〗 is 5 , R (5, π), 7, 々-20 -amino ten, hydrazone} 〇 丨 dimethyl_3,, 5 丨 _dioxospiro [furan_2 (3Η), 17 '" (5'11)-[7,4] methylenefluorene [411] cyclofluorene U] phenanthrene] nitrile. In the next step of the synthesis of Scheme 1, the enamine of formula νπ is hydrolyzed to produce a dione compound of formula VI (Please read the notes on the back before filling in this page j

VI-Ordered by the Central Bureau of Standards of the Ministry of Economic Affairs, Consumer Cooperatives, where -A-A-, R3, R8 and R9 are as defined in the formula ¥ 111. Trihydrolysis can use any water-containing organic or inorganic acid. Preferred is argon acid. To promote productivity, it is best to use organic solvents that are miscible with water, such as low carbon alkanols, as co-solvents. The acid should be present in a ratio of νπ and at least one equivalent of the enzyme. In an aqueous system, at approximately 80 ° C, the amidoamine enzymatic substance νπ can be converted to formula VII g in approximately $ hours. Operating at elevated temperatures increases productivity, but temperatures 67-

593305 A7 B7 is called Y ^ YCCH2] 7

V. Description of the invention (65 is not decisive. Determine the appropriate temperature based on the solvent system and acid volatility.

The preferred enamines of formula VII are enzymatically compatible with formula vIIA • czrx

VIIA

And the diketone product conforms to the formula VIA (please read the precautions on the back before filling this page) H0 / v ^ Pt, CCH2] "= x

, 1T VIA X are printed in the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs in the formula VIIIA, where the parent _ Α · Α_, _B-B-, Y1, γ2 and the definitions. At the end of the reaction period, the solution was cooled to zero. 〇 and 25 ^, so that the product: crystal. The crystallization of the product can be reconfirmed from a suitable solvent such as isopropanol or methanol in order to produce a product suitable for use in the next step of the process, but recrystallization is generally not required. The compound of formula VI is a novel compound, and it has the compound of formula I, especially the compound of formula I-68- This paper size is suitable for household standard (CNS) A4 specification (21Qx 297 male thin 593305 A7 B7 V. Description of the invention (66) <the actual valence of the intermediate. The preferred compound of formula VI conforms to the formula α ^ -β_βκΗ2.2_, with hydrogen, low carbon number ㈣ or low carbon number alkoxy group, and R8 and R9 together. Form 20_spiroxane ring:

The Central Consumers Bureau of the Ministry of Economic Affairs's Consumer Cooperatives printed XXXIII-style VIA. The most preferred compound of formula VI is 4, s (4, a), 7, α-hexadecane-11-hydroxyl-10f 羟基 -13 丨 /? _ Dimethyl_3 丨, 5,20, _Trioxospiro [furan 2 (3H), 17 0-[4,7] methyl [17H] cyclopenta [a] phenanthrene] -5, 10,000 (2! 11) &quot; In a particularly preferred embodiment of the present invention, the method described above produces a sulfonamide of formula VII from a compound of formula V111 and converts it to a diketone of formula VI in situ. In this specific embodiment of the present invention, as described above, the enzyme of formula VIII is reacted with an excess of an alkali metal cyanide in an aqueous solvent containing a proton source, or in the presence of a base and an alkali if necessary, Reacts with excess ketocyanohydrin. However, instead of cooling the reaction mixture, acidify and add water in a calculated proportion so as to cause precipitation of the enamine, and it is best to avoid actual cooling of the reaction mixture. At the end of the cyanation reaction, water and an acid, preferably an inorganic acid such as sulfuric acid, are indeed added to the mixture, and the proportion of the acid added is sufficient to neutralize the excess alkali metal cyanide, usually per mole. Ear-type VIII enzymes require the introduction of at least one mole equivalent of acid, preferably between about 2 and about 5 moles of enzyme per equivalent. However, it is necessary to maintain a high temperature and a high degree of dilution sufficient to avoid actual precipitation and to allow enamine to diketone hydrolysis in the liquid phase. Therefore, the method is performed with minimal interference and high productivity. The best hydrolysis effect (please read the precautions on the back before filling this page) Clothing · Order-69 · This paper size is applicable to China National Standard (CNS) A4 specification (210X 297 mm) 593305 A7 B7 V. Description of the invention (67 ) Is operated at a temperature of at least 80 ° C, more preferably at about 90 ° to about 100 ° C, usually for about 1 to about 丨 0 hours, preferably about 2 to about 5 hours . The reaction mixture is then cooled, preferably at a temperature between about 0 ° C and about 15 ° C, advantageously in an ice bath, to about 5 ° C to about 10 ° C, so that the formula v The diketone product of I precipitates. Solid products can be recovered by filtration, and impurities can be removed by rinsing with water. In the next step of the synthesis of plan 1, the diketone compound of formula VI is reacted with a metal alkoxide, the ketone bridge between the 4 and 7 positions is opened, and the bond between the carbonyl group and the 4-carbon is cut to form Substituted oxycarbonyl substituent at the 7-position and excludes cyanide on the 5-carbon. The product of this reaction is an acyl ester compound conforming to formula V (please read the precautions on the back before filling this page)

-· & Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economics where -A-A-, R3, -B-B-, R8 and R9 are as defined in formula viii, and R1 is a low-carbon oxycarbyl or carbonyl group. The metal alkoxide used in this reaction conforms to the formula R1GOM, where M is an alkali metal and rig is equivalent to the alkoxy substituent of Ri. When the metal alkoxide is potassium methoxide or sodium, the production capacity of the reaction is best satisfied, but other low carbon number alkoxides may be used. The alkoxide is particularly good. Phenates, other aryl oxides, and aryl sulfides can also be used. This reaction is conveniently carried out in the presence of an alcohol conforming to the formula R1GOH, where R1G is as defined above. Other traditional solvents can be used. Preferably, the enzymatic substance of formula VI is between about 2% by weight and about 12% by weight. -70- This paper size applies to China National Standard (CNS) A4 (210X 297 mm) 593305 Central Standard of the Ministry of Economic Affairs A7 B7 printed by the Bureau ’s Consumer Cooperatives V. Description of the invention (68 ratios exist, more preferably at least about 6% by weight, and R 丨 〇M is negatively affected by about 0.5 to about 4 moles per mole The proportion exists. The temperature is not strictly limited, but the elevated temperature can promote productivity. The reaction time is usually between about 4 to about 24 hours, preferably about 4 to 16 hours. Depending on the solvent used, the reaction can be Conveniently, it is done at the reflux temperature of atmospheric pressure. In the conversion of the diketone of formula VI to the hydroxy ester of formula VI, the ion of the byproduct cyanide may react with the product to form 5-cyanoester. Because at low concentrations It is better for this equilibrium, so it is best to carry out the reaction at a high dilution, for example, for the reaction with sodium methoxide, a high dilution of 40: 1. It has been found that using potassium methoxide instead of sodium methoxide can obtain Markedly Because potassium methoxide is the reagent, a dilution in the range of about 20: 1 is usually sufficient to minimize the degree of reverse cyanation. According to the present invention, it has been further disclosed that By using appropriate chemical or physical measurements, the by-product cyanide ion is removed from the reaction zone to suppress the reverse cyanation reaction. Therefore, in a further and further embodiment of the present invention, the dione and the genus The alkoxide reaction can be performed in the presence of a reagent that causes cyanide ions to form, such as, for example, salts including cations that form insoluble cyanide compounds. Such salts may include, for example, zinc zinc, iron sulfate, Or alkaline earth metals or transition metals are substantially more soluble than the corresponding cyanides, any salts, sulfates or other salts. If the decomposed words are present in a ratio of about one equivalent of the enzymatic substance per equivalent of the diketone, Compared to operating this method in the absence of alkali metal dentate, a substantial increase in the productivity of this reaction has been observed. &Quot; Even if a precipitant is used to remove cyanide ions, it is best to Quite high (please read the notes on the back before filling this page)

-71- 593305 A7 B7 V. Description of the invention (69)

The 'mole ratio of solvent to diketone substrate' can be significantly reduced by the use of a precipitant, compared to the reaction lacking such a reagent. Recovery of the hydroxyl group of formula V can be accomplished according to the extraction or non-extraction procedures described below. The preferred dione of formula v I is enzymatically compatible with formula V 丨 A

And the ester product conforms to formula VA

VIA

(Please read the notes on the back before filling this page)

Employees of the Central Standards Bureau of the Ministry of Economic Affairs, Consumer Cooperatives, and R1 are as defined in Formula V. The product of Formula V is a novel compound that has the actual price of preparing a compound of Formula I, especially an intermediate in Formula IA. A preferred compound of formula V is in which B-B- is _CH2-CH2-, R3 is hydrogen, low-carbon alkyl or low-carbon alkoxy, and R8 and R9 together form 2_spiro Oxane ring: XXXI Engineering-72- This paper size is applicable to Chinese National Standard (CNS) A4 (210X 297 mm) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 593305 A7 B7 V. Description of the invention (70) VA compound. It is preferred that the compound of formula V is n-, 17 "-diazonyl-3 -oxoprogestin-4-fluorene-7α, 21-dimethanoate, r-lactone. By using, for example, concentrated HC 1, acidify the reaction solution, cool to ambient temperature, and extract the product with an organic solvent such as dichloromethane or ethyl acetate to isolate the compound of formula V. rinse the extract with an aqueous alkaline rinse solution, and dehydrate And filtered, followed by removal of the solvent. Alternatively, the reaction solution containing the product of formula V can also be stopped with concentrated acid. The product solution is concentrated, cooled to 0 ° C to 25 ° C 'and the product solids are isolated by filtration According to a better mode for recovering the product of formula V, after the reaction period, methanol and HCN are removed by steaming, and water and acid are added before or during the distillation. The operation can be simplified if water is added in the steam room. However, during the steaming period, the volume of the guest in the steel of the steaming hall is actually kept constant. During the distillation process, the product of formula V forms crystals from the bottom of the distillation pot. This recovery mode provides no extraction required Handled high quality crystalline product. According to another option, after removing the solvent by steaming the crane, the reaction solution containing the product of formula V can be stopped using an inorganic acid, such as 4N HC1. The removal of the solvent is for removing the residual H c N from the reaction product. It may also be effective. It has been found that multiple solvent extraction is not necessary for the purification of compounds of formula V. The compounds of formula V act as intermediates in the preparation of epoxy mexenos, as in this article. In fact, this type of extraction is usually completely excluded. Solvent extraction is used for the purification of the product, and solvent washing and alkaline washing with brine are wanted. However, where solvent extraction is excluded, it is also excluded. Salt water and alkaline washing. Excluding extraction and washing significantly increased the productivity of this method, and will not yield-(Please read the notes on the back before filling this page), 1Τ -73 593305 Staff of the Central Standards Bureau of the Ministry of Economic Affairs Printed by the consumer cooperative V. Description of the invention (71 A7 B7 or the quality of the product, it also excludes the need for T # 丄, λ to dehydrate the washed solution: :: such as sulfuric acid Desiccants such as hydroxy ~~ alkaneoxy :: next step in the method, which will re-dissolve the crude t at the u position in the solvent formula compound: Good release

IV,, -A-A_, R, -B_B_, R8 and R9 are as defined in formula X1π, R is as synonymous in formula V, and R2 is a low carbon number aryl group. The radicals are fluorenyl, alkoxy, or autogenous. Preferably, the 11-hydroxyl group is esterified by adding a low-carbon alkylsulfonyl sulfonate, fluorenyl radical or acid anhydride to a solution containing an intermediate product of formula V and reacting therewith. Low-carbon alkylsulfonyl sulfonates are preferred, especially methanesulfonyl sulfonium. In addition, by reacting with an appropriate reagent, such as thionyl bromide, thionyl chloride, sulfonyl chloride, or grass gas', the 1 1 α _ radical can be converted into a halide. Other reagents capable of forming 1 1 slave-sulfonate include tosylsulfonium chloride, benzenesulfonium sulfonium gas, and trifluoromethanesulfonic anhydride. The reaction is run in a solvent containing a hydrogen scavenger such as triethylamine or pyridine. Inorganic tests such as carbonate or sodium can also be used. The starting concentration of the light ester of formula V is preferably between about 5 wt% and about 50 wt%. The esterification agent is preferably in a slight excess. Dichloromethane is a particularly suitable reagent for this reaction, but other solvents can also be used, such as digas ethane, pyridine, chloroform, fluorenyl ethyl ketone, dimethoxyethane, methyl isobutyl-74. China National Standard (CNS) A4 specification (210X297 mm) (Please read the precautions on the back before filling this page) Clothing · 593305 A7 B7 V. Description of the invention (72) ^-Ketones, acetone, other ketones, ethers Class, acetonitrile, toluene and tetrahydrofuran. The reaction temperature is mainly determined by the volatility of the solvent. In dichloromethane, the reaction temperature is preferably between about -10 ° C and about 10 ° C. The preferred light-based ester of formula V is enzymatically compatible with the formula va rwJ η

C two X

VA

And the product conforms to formula IVA

〔Ch2〕 · ^ ~~ G—X (Please read the precautions on the back before filling in this page)

IVA wherein _A_A_, _B_B_, γΐ, Y2 and x are as defined in Formula XIII, R1 is a lower alkoxycarbonyl or hydroxycarbonyl group, and R2 is as defined in Formula IV. The product of Formula V printed by the Employees 'Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs is a novel compound which has a practical value as an intermediate for the preparation of a compound of Formula I', especially of Formula IA. Preferred compounds of formula iv, in which -A_A- and -B_B · are -CH2-CH2-, R3 is hydrogen, lower alkyl or lower alkoxy, and R8 and R9 together form 20-spiro Oxane ring:

XXXIII -75- Exact, and, scales are applicable to Chinese National Standards (CNS) A4 specifications (210X 297 mm 593305) Consumers Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs m Α7 Β7 V. Compounds of formula (IVA) of the invention description (73). Preferably, the compound of formula IV is 17 "_hydroxy_η π-(methazinyl) oxy-3-oxoprogesterone_4_ene-7 α, 2ΐ_diI hydrogen acid methyl ester, r -lactone If desired, the compound of formula V can be separated by removing the solvent. Preferably, the reaction solution is first rinsed with an aqueous rinsing solution, such as 0.5-2N NaOH, and then with Acidic rinsing solution, such as 0-5 · 2N He 丨. After removing the reaction solvent, for example, by dissolving the product in difluoromethane: k, and then adding other solvents such as ethyl The solubility of the product of formula ν causes the product to precipitate in crystalline form and cause it to recrystallize. In the recovery of the product of formula IV or in the preparation of the reaction solution, as described below, 'in order to intermediate formula IV Compounds are converted to intermediates of formula π, if the ion-exchange resin is used instead to treat the solvent In order to remove acidic and alkaline impurities, all extraction and / or rinsing steps can be eliminated. The solution is first treated with an anion exchange resin and then treated with a cation exchange resin. In addition, an inorganic adsorbent can be used first Treatment of a reaction solution, such as basic alumina or basic silica, followed by rinsing with dilute acid. Basic silicon dioxide or basic alumina, usually at a ratio of between about 5 and about 50 grams per kilogram of product, It is mixed with the reaction solution, preferably between about 15 and about 20 grams per kilogram of product. Regardless of whether an ion exchange resin or an inorganic adsorbent is used, the resin or the inorganic adsorbent can be used to surround the reaction solution together with the reaction solution. The slurry is simply formed by stirring at temperature, and then the resin or inorganic adsorbent is removed by filtration to complete the treatment. In another preferred embodiment of the present invention, a portion is removed by removing The paper size is applicable to the national standard (CNS) A4 specification of the towel (21〇 × 297 mm) '— ~~. (Please read the precautions on the back before filling this page) Order 593305 A7 ---- -_B7 __Fifth, the formulation of the invention (74), the concentrated solution of the compound of the formula IV is recovered in a crude form. The concentrated solution is used directly in the next step of the method, from the compound of the formula IV Removal of the n "-releasing group, thereby generating a phosphonium ester of the formula π:

(Please read the precautions on the back before filling out this page) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs. Α-Α_, R3, _Β_Β_, $ 8 and Hua 9 are as defined in the formula ¥ 111, and R is as in Definition in Formula V. For the purpose of this reaction, the R2 substituent of the compound of Formula 1 may be any releasing group which is effective to generate a double bond between 9 and 1 carbon &lt; when extracted. It is preferred that the releasing group is a low-carbon alkylsulfonyloxy or fluorenyl substituent, which is removed by reaction with an acid and an alkali metal salt. Inorganic acids can be used, but low carbon number alkanoic acids are preferred. Advantageously, the reagents related to the reaction further include alkali metal salts of the chain burning acid used. Particularly preferably, the releasing group includes methanesulfonyloxy, and the reagents related to the reaction include formic acid or acetic acid, and alkali metal salts of one of these two acids or other low-carbon alkanoic acids. When the releasing group is methanesulfonyloxy group and the removal reagents are formic acid and potassium formate, a higher proportion of 9, i i to 11,12-alkene is observed. If free water is present during the removal of the free radicals, it is easy to form insoluble materials, especially 7,9-lactone -77- This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm)

1T 593305 Α7 Β7 V. Description of the invention (75)

It is not easily removed from the end product. Therefore, acetic anhydride or other drying agents are used to remove the water present in the formic acid. Measured by Karl Fischer's analysis of water, based on the total reaction solution before the reaction, the free moisture content in the reaction mixture should be maintained below about 0.5% by weight, and preferably below about 0.1% by weight. Although the reaction mixture is preferably maintained at a dryness of 0.3% by weight of water, where practical and satisfactory results have been obtained. Preferably, the reaction charge mixture contains from about 4% to about 50% by weight of an enzyme of formula Iv in an alkanoic acid. Preferably the alkali metal salt of the acid is included at about 4% to about 20% by weight. When acetic anhydride is used as the desiccant, it is preferably present at a ratio of from about 0.05 mole to about 0.2 mole per mole of alkanoic acid. It has been found that the proportion of by-products 7,9-lactone and 1 1,1 2-alkene in the reaction mixture is quite low in excluding reagents containing a combination of trifluoroacetic acid, trifluoroacetic anhydride and potassium acetate, This reagent is used as a reagent which excludes the release group and forms an ene ester (9,1 1 · chain fluorene). Trifluoroacetic anhydride is used as a desiccant, and based on the trifluoroacetic acid exclusion agent, it should be present in a proportion of at least about 3% by weight, more preferably at least about 15% by weight, and most preferably about 20% by weight . In addition, 11 free radicals can also be excluded from the compound of formula IV. By heating -78- This paper size applies Chinese National Standard (CNS) A4 specification (210X 297 mm) (Please read the precautions on the back before filling in this Page), printed by the Consumers' Cooperative of the Central Standards Bureau, Ministry of Economic Affairs, 593305 A7 B7 5. Description of the invention (76) The formula in an organic solvent such as DM SO, DMF, or DMA produces the purpose of formula 11. Still further according to the present invention, a compound of formula IV is initially made with an alkanoate such as isopropenyl acetate in the presence of an acid such as toluenesulfonic acid, or an anhydrous inorganic acid such as osmic acid React to form a 3-enol ester of a compound of formula V:

(Please read the precautions on the back before filling in this page.) V (z) In addition, you can also use acid anhydride and alkali to treat, such as acetic acid and sodium acetate, to form 3-enol ester. In addition, it may include treatment with ketene in the presence of an acid to generate IV (Z). The intermediate is then treated with an alkali metal formate or acetate in the presence of formic acid or acetic acid, formula IV (Z), to yield Δ-9,1 1 1-alcohol acetate of formula IV (Y): Central Ministry of Economic Affairs Standards Bureau, Consumer Cooperatives, India

IV (Y) can then be converted into the formula in an organic solvent, preferably an alcohol such as methanol, through the thermal decomposition of enol ethyl, or its reaction with a metal alkoxide. 11 esters. The exclusion reaction is highly selective for the esters of formula I 丨 -79- This paper size applies Chinese National Standard (CNS) A4 (210X297 mm) 593305 A7 B7 V. Description of the invention (77)

The selectivity is most preferred, and this selectivity can be preserved in the conversion of i 2 _ olefins and 7: esters to fluorenone. The better formula IV is subject to enzymes, which matches the formula! v A -lactone, and ethanol

IVA (Please read the notes on the back before filling in this page j

CZZX Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

And the objective product meets formula 11 A

IIA where -Α_Α_, -Β_Β_, γΐ, Y2, and X are respectively defined as in the formula χιπ, and R1 is defined as in the formula V. If desired, the compound can be isolated by removing the solvent, dissolving the solid product in cold water, and extracting it with an organic solvent, such as ethyl acetate. After proper rinsing and dehydration steps, the product is recovered by removing the extraction solvent. The fluorenyl ester is then dissolved in a solvent suitable for converting the product of formula I. Alternatively, it is also possible to preferentially remove the 7,9-lactone by adding water to the concentrated product solution and filtering the solid product to isolate the ethyl ester. The enzyme-80 of formula 11 is adapted to the Chinese national standard (CNS) A4 size (210X 297 mm) 593305 A7 B7 Five invention descriptions (78 The printed matter of the staff consumer cooperative of the Central Standards Bureau of the Ministry of Economic Affairs is transformed into formula IA The product can be operated by the method interpolated in U.S. Patent No. 4,559,332, which is hereby incorporated by reference in particular, or, more specifically, by a novel reaction, using an amidated acetamide starter, as follows In other specific embodiments of the present invention, a hydroxy ester of formula can be converted to a phosphonium ester of formula II without the need to separate the intermediate of the compound of formula 1 ¥. In this method, the hydroxy ester is dissolved In an organic solvent, such as dichloromethane; then add a hydrating agent, such as methanesulfonyl chloride, or a sulfonating agent, such as sulfonium gas, to the solution. Stir the mixture, and if a dentifying agent is involved, then L HC1 scavengers such as imidazole I. Mixing of the base with the solution is highly latent and should therefore be operated at a controlled rate and cooled sufficiently: After the addition of the base, the resulting Mix The material is allowed to warm to an appropriate temperature, for example, 0 C to 罜, or slightly exceeded, and the reaction is continued for a period of time, usually 1 to 4 hours. After the reaction is completed, the solvent is stripped, preferably _ 丨 to + 15 At a high altitude (eg, 24 "to 228, Hg), it is more preferably about 0 ° C to about 5 ° C in order to concentrate the solution and remove excess alkali. The enzyme is then redissolved in Among organic solvents, halogenated solvents such as dichloromethane are preferred for conversion to vinyl acetate. The ion-exclusion reagent is preferably prepared by mixing organic acids, organic acid salts, and desiccants in an anhydrous reactor. For the preparation, the preferred ones are formic acid, alkali metal formate and acetic anhydride. The addition of acetic anhydride is exothermic and results in the release of CO, so the rate of addition must be controlled. To promote the removal of water The temperature of the reaction is preferably maintained in the range of ⑽ ^ to ... ^ ', preferably about 65 ° C to about 75 ° c. Then add this reagent to the formula (Please read the note on the back first (Please fill in this page again for details) Order 81- Staff Consumption of Central Bureau of Standards, Ministry of Economic Affairs Printed by Suzakusha 593305 A7 ________B7 V. Description of the invention (79) In the product solution of t to complete the elimination reaction. After 8 hours of drawing, it is best to heat the reaction mixture to a temperature of at least about 8 5 r, but not Over approximately 95 ° C until all volatile distillates have been removed, and then continue for an additional period of time to complete the reaction, typically 1-4 hours. The reaction mixture is cooled and subsequently recovered by standard extraction techniques. I want to recover 晞 g by evaporating the solvent. I also found that by using another procedure that does not require an extraction step after the reaction is eliminated, the enyl ester of formula j can be recovered from the reaction and valley liquid, thereby providing a cost Savings, yield improvements and / or productivity improvements. In this method, the ene ester product is precipitated by diluting the reaction mixture with water after removing formic acid. The product was then isolated by filtration. No extraction is required. According to another option, the hydroxy ester of formula v is converted to the ester of formula π, without isolating the compound of formula IV, replacing the ii "-hydroxy group of the hydroxy ester of formula V with hydrogen, and then dehydrogenating by heat _Chemical formation is in situ; the enyl ester of ^. Substitute the substitution of several groups with hydrogen, and by sulfonation with, it is better to continue hydrazine 'in the cold, such as in imidazole. The reaction is completed in the presence of a hydrogenation scavenger. The hydroxyester is dissolved in a solvent such as tetrahydrofuran and cooled to 0 ° C to -7 ° C. Sulfonation is added and the reaction mixture is warmed. To an appropriate temperature, such as room temperature, for a period of time sufficient to complete the elimination reaction 'is usually 1 to 4 hours. The method of this specific example does not merely mix two steps into one, but also excludes the use of the following substances: Solvents (such as acetic acid); and desiccants (acetic anhydride or sodium sulfate). In addition, the reaction does not require reflux conditions, and avoids the production of by-products C 0 when using acetic acid as a desiccant.- 82- This paper size applies to China Standard (CNS) A4 (21 0x297 mm)-(Please read the notes on the back before filling out this page) Order ^ 1. Staff Consumer Cooperatives, Central Standards Bureau, Ministry of Economic Affairs, India Fan 5933305 5. Description of Invention (8 〇) According to a particularly preferred embodiment of the present invention, the dione compound of the formula 1 ¥ can be converted to epoxy mexanol or other compounds of the formula, without the need to isolate any intermediates in pure form. According to this preferred The method uses a strong acid solution to stop the reaction solution containing the hydroxyester, cools to the surrounding temperature, and then extracts it with an appropriate extraction solvent. Advantageously, before the extraction, an aqueous solution of an inorganic salt is added to the reaction mixture, for example, 0% by weight of a physiological saline solution. The extract is rinsed and dehydrated by azeotropic distillation to remove the methanol solvent remaining in the ketone cleaving reaction. Then use a compound of formula v at about 5% to about 50% by weight The resulting concentrated solution is contacted with a sulfonating agent or an alkylating agent under cold conditions to form a sulfonic acid ester or a dicarboxylic acid ester. After the alkyl sulfonation is completed, After the carboxylation reaction, the reaction solution is passed through a column covered with an acidic and then a basic fork-changing resin in order to remove basic and acidic impurities. After passing through each column separately, rinse the column with a suitable solvent, such as two Chloromethane in order to recover the residual sulfonic acid or dicarboxylic acid esters from it. The mixed eluate and rinse fractions are mixed and preferably reduced under vacuum to produce Sg or a few &amp; S concentrated solution. The concentrated solution is then contacted with a desiccant including a reagent that causes removal of the 1 1 α-ester release group and removes hydrogen from the 9, u _ double bond. The release group is removed The reagent preferably includes the formic acid / metal formate / acetic anhydride desiccant solution described above. After the reaction is completed, the reaction mixture is cooled and the formic acid and / or other volatile components are removed under air. The residue is cooled to ambient temperature, subjected to a suitable rinsing step, and then dehydrated to obtain a concentrated solution containing a formula II ester. The olefinic ester can then be converted to -83 using the method described in this article or in US Patent No. 4,559,332. This paper scale is applicable to the Chinese National Standard (CNS) A4 specification (210X 297 mm) (Please read first Note on the back, please fill in this page again), 11 -I «^ 1— — ^ 1 Printed by the Employee Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 593305 A7 I ---------- B7____ V. Description of the Invention (S1 ) Epoxy Mexanol or other compounds of formula I. In a particularly preferred embodiment of the present invention, the solvent is removed from the reaction solution under airspace and the product of formula IV is distributed between water and a suitable organic solvent 'such as ethyl acetate. The back extract is then washed with an organic solvent and an extract layer, and the solution is preferably an alkali metal hydroxide solution containing an alkali metal dentate. The organic phase is concentrated, preferably under argon, to produce the enyl product of formula II. The product of formula η can then be dissolved in an organic solvent, such as digas methane, and further reacted in the manner described in the Patent No. 332 to produce the product of formula I. When tri-acetonitrile is used in epoxidation, it has been found that the choice of solvent is important. The tritiated solvent is preferred, and dichloromethane is particularly preferred. Solvents such as dichloroethane and gaseous benzene yield fairly satisfactory yields, but generally yield better yields in dichloromethane reaction media. Solvents such as acetonitrile and ethyl acetate usually result in poor yields, while reactions in solvents such as methanol or water / tetrahydrofuran give a fraction of the desired yield. Still further according to the present invention, it has been revealed that by using diethylamidamine instead of di-acetonitrile as a peroxide activator with respect to the epoxidation reaction, many improvements can be obtained in the synthesis of epoxy mexanol. According to the special method, the epoxidation is completed by reacting the enzyme of formula IIA with hydrogen peroxide in the presence of trichloroacetamide and a suitable buffer solution. Preferably, the reaction is carried out at a pH range of about 3 to about 7; preferably, about 5 to about 7. Regardless of these factors, however, successful reactions have been achieved outside the preferred pH range. -84- This paper size applies to China National Standard (CNS) A4 (210x297 mm) —- (Please read the precautions on the back before filling this page}

Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 593305 M —____ B7 V. Description of the Invention (82) Use a buffer solution composed of dipotassium hydrogen phosphate, and / or use a solution between approximately 1: 4 and approximately 2: 1 The relative ratio, preferably in the range of about 2: 3, is particularly favorable with a buffer solution composed of dipotassium hydrogen acetate and potassium dihydrogen acetate. Boric acid buffer solutions can also be used, but usually result in slower conversions than dipotassium phosphate or KAPO4 or K: 2QPO4 / KQ2P04 mixtures. Regardless of the composition of the buffer solution, the pH range indicated above should be provided. Regardless of the overall composition of the buffer solution or the actual pH it may give, it has been observed that if at least a portion of the buffer solution is composed of a binary hydrogen phosphate ion. The response will be much more efficient. Xian believes that in the formation of addition compounds or complexes that include initiators and hydroperoxide ions, they essentially share the role of homogeneous catalysts, and their alternating production may react to the entire epoxidation reaction mechanism. Words are necessary. Therefore, the quantitative requirement for the dibasic hydrogen phosphate (preferably from k2hP04) may be only a small amount of catalytic concentration. In general, it is preferred that HPO4 is present in a ratio of at least about 0_i equivalents per equivalent of enzyme, such as between about 0.1 to about 0.3 equivalents. The reaction is carried out in a suitable solvent, preferably digas methane, but other solvents such as chlorobenzene or dichloroethane can also be used. It was also found that toluene and mixtures of toluene and acetonitrile were also satisfactory. Needless to say that the theory is sufficient, it is assumed that the reaction proceeds most efficiently in a two-phase system, in which a hydroperoxide intermediate is formed and distributed to the organic phase with low water content, and then reacts with the organic phase in the organic phase. Subject to enzymatic reactions. Therefore, preferred solvents are those with lower water solubility. By including other agents such as acetonitrile, more efficient recovery from toluene can be facilitated. Yin ----------- (Please read the notes on the back before filling this page)

, 1T

593305 A7 B7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (83 In converting the enzyme of formula 11 to the product of formula I, toluene provides an advantageous process because the enzyme is freely dissolved in toluene And the product is not. So 'the product precipitates out when the transformation reaches the range of 40-50% during the reaction' produces a three-phase mixture from which the product can be conveniently separated by filtration. In this method In the conversion step, methanol, ethyl acetate, acetonitrile alone, THF, and THF / water cannot provide such effective results as halogenated solvents or toluene. Digas is the favorite reagent, but other three Halogenated acetamide, such as difluoroacetamide. Trimethylated methylbenzamide can also be used, as well as other aryl groups between the electron-withdrawing tri-automated methyl group and the carbonyl group of fluorenamide. Compounds based on phenols. 3,3,3_trihalogenopropylammonium amine can also be used, but with poorer results. Generally peroxide activators can fit the formula: R ° C (〇) NH2 where R ° For having at least something like monochloride Group with the same high electron-withdrawing strength (measured by the sigma constant). More preferably, the peroxide activator can meet the following formula: f 〇X —C—Rp-c! — Where X, χ and χ3 are selected from halogen, hydrogen, alkynyl, dentyl, chloro and cyanoalkyl, and rp is selected from arylene and _ (cx4x5) n_, where at least one of XXXX and X is halogen Or perhaloalkyl. When any one of X1, X2, X3, X4, or X5 is not a prime, it is preferably a self-fired group, most preferably a full-line group. Particularly preferred activators include n (please (Read the notes on the back and fill out this page)

-86 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 593305 A7 ____B7 V. The description of the invention (84) is 0, and at least two of X1, X2, and X3 are halogen; or where X1, X2, X3, X4, and X5 are all Those are all halogen or perhalogenated. Preferably, χΐ, X2, χ3, χ4, and χ5 are C1 or F, respectively, and most preferably C1, although mixed compounds are also suitable, such as perchloroalkyl or perbromoalkyl and mixtures thereof. Preferably, the peroxide activator is present at a ratio of at least about 1 equivalent per enzyme equivalent of the enzymatic substance initially present, and more preferably between about 5 and about 2 equivalents. The reaction should be loaded with at least a slight excess of hydrogen peroxide or gradually added as the ring emulsification proceeds. Although only one to two equivalents of hydrogen peroxide are consumed per mole of the enzyme in the reaction, it is preferred that the hydrogen peroxide be loaded in an amount that actually exceeds the amount of enzyme and activator originally present. Without limiting the present invention by a specific theory, I believe that the reaction mechanism involves the formation of an addition compound of activator and 〇 ，, the formation of the reaction is reversible, and the equilibrium is favorable for the reverse reaction, so in order to make the reaction proceed forward, Requires an initial practical excess of hydrogen peroxide. The temperature of the reaction is not strictly limited, and it can be efficiently completed in the range of 0 ° C to 100 ° C. The most appropriate temperature depends on the solvent selected. Generally the preferred temperature is between about 20 and 30, but in some solvents, such as toluene, the reaction can advantageously operate in the range of 600-7 (rc.) At 25 ° C, the reaction It usually takes less than 0 hours, usually 3 to 6 hours. If additional activators and hydrogen peroxide are needed, they can be added at the end of the reaction cycle in order to achieve the complete conversion of the enzyme. At the end of the reaction cycle, remove In addition to the liquid phase, it is best to rinse the organic reaction solution 'in order to remove water-soluble impurities, and then the product can be recovered by removing the solvent. Before removing the solvent, the reaction solution should be rinsed and used at least (please read first (Notes on the back then fill out this page)

1T -87- 593305 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs

A7 B7 5. Description of the invention (85) M and a moderately sensible rinsing solution, such as sodium carbonate. The reaction mixture is preferably continuously rinsed with a mild reducing solution, such as a weak (eg, 3% by weight) aqueous sodium sulfite solution; an alkaline solution, such as NaOH or KOH (preferably about 0.5N ); An acidic solution, such as HC1 (preferably about 1 N); and a final neutral rinse, including water or saline, preferably saturated saline 'in order to reduce product loss. Prior to the removal of the reaction solvent, other solvents, such as organic solvents, preferably ethanol 'can be advantageously added so that after more volatile reaction solvents are removed by distillation, the product can be recovered by crystallization. It should be understood that novel epoxidation methods using dichloroacetamide or other novel peroxide activators have applications that far exceed the various plans for the preparation of epoxy mexanol, and in fact can be used in a variety of ways in the liquid phase. The olefinic double bond of the enzymatic substance undergoing the reaction is used for the formation of epoxide. This reaction is particularly effective for unsaturated compounds in which the olefinic carbon is tetra- and tri-substituted, that is, RaRbC = CRcRd and RaRbc = CRCRH, where ^ to ... represent substituents other than hydrogen. When the substrate is a cyclic compound with a triple-substituted double bond, or a cyclic or acyclic compound with a tetra-substituted double bond, the reaction proceeds most rapidly and completely. Representative enzymes of this reaction include △ _9,1 1 -Conoreno, and

-88-This paper size is applicable to China National Standard (CNS) A4 (210X 297mm) (Please read the precautions on the back before filling this page) Order 593305 A7 B7

〇 Because the reaction proceeds more quickly and completely with double bonds with tri- and tetra-substitutions, in compounds that may contain other double bonds where the alkenyl carbon is mono- or even di-substituted, spanning such double bonds The selective epoxidation is particularly effective. It should be further understood that this reaction can be advantageously used in the epoxidation of mono- or even disubstituted double bonds, such as 11,12-alkenes in various steroid receptors. However, because the highly substituted double bonds, such as 9,1 1 -strands, are preferentially epoxidized and have high selectivity, the method of the present invention achieves the epoxidation step for various reaction schemes described elsewhere herein. High yields and productivity are particularly effective. Improved methods have been shown to work well with the following:

This paper size applies to Chinese National Standard (CNS) A4 specification (210 × 297 mm) (Please read the precautions on the back before filling this page)

〇 Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economics Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economics 593305 A7 -------- B7 V. Description of Invention (87)

It is a particularly advantageous application. Multiple advantages have been demonstrated for the method of the present invention, in which trigas acetamide is used instead of trigas acetonitrile as a reagent for transferring oxygen in the epoxidation reaction. For the epoxidation of acetamide, the reagent system for epoxidation across tri-substituted double bonds, using the double substitution in the same molecular structure and "0_ketoalkenes" provides strict regional control. Therefore, it actually increases The reaction yield, product profile, and final purity were further revealed. It was further revealed that triacetam did not experience the actual excess oxygen production observed with tris-acetonitrile, giving improved safety to epoxidation. And Sangas In contrast, acetonitrile promotes the reaction. The reaction of acetamide shows the least exothermic effect, so it is beneficial to control the thermal profile of the reaction. Observe that the effect of agitation is minimal and the reactor efficiency is more consistent, and there are more wins. Advantages of the three-gas acetonitrile method. This reaction is more expensive than the reaction promoted by dichloroacetonitrile and can be scaled up. The isolation and purification of the product is simple, and I have not experienced anything like using m-chloroperoxybenzoic acid or other Significant carbonyl function encountered in peroxyacid Bayer-Willieg-90- This paper size is applicable to China National Standard (CNS) A4 (210X297 mm) (Please read first Note the surface to fill out this page}

593305 A7 B7 V. Description of the invention (88) (Bayer-Villager) oxidation (peroxide promotes the conversion of ketones to vinegar), and the reagent is cheap, easy to obtain and easy to operate. The novel epoxidation method of the present invention is very useful in the inference step of the synthesis of plan 1. In a particularly preferred embodiment, the complete method of the Plan 1 process is as follows:.

(Please read the notes on the back before filling this page)

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-91-

This paper size applies to China National Standard (CNS) A4 (210X 297 mm) 593305 A7 B7 · 5

2. Description of the invention (89 The second novel reaction plan (Plan 2) of the present invention, starting from Conorno or other enzymes conforming to formula XIII)

XIII. In the first step of the method, the enzymatic substance of formula XII is converted to the product of formula XII using a cyanation reaction scheme substantially the same as that described above for converting the enzyme of formula VIII to an intermediate of formula VII (Read the notes on the back before filling out this page)

Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs

XII

It is preferred that the enzymatic substance of formula XIII conforms to formula XI J A y 2 c = z: x and the enamine product conforms to formula XΙΙΑ-92- This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm)

XIIIA 593305 A7 B7 V. Description of the invention (90

XIIA where-A-A-, -B-B-, Y1, Y2, and X are as defined in Formula XIII, respectively. In the second step of Scheme 2, the enamine of formula XII is hydrolyzed to the intermediate diketone product of formula XI using a reaction scheme substantially the same as that described above for converting the enzyme of formula VI11 to the intermediate of formula VII.

(Please read the notes on the back before filling this page)

XI Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs

Among them,-AA-, R3, -B_B-, R8, and R9 are as defined in formula VIII. It is better that the enzymatic substance of formula XII complies with formula XIIA-93- This paper standard applies to China National Standard (CNS) A4 (210X 297 mm) 593305 A7 B7 V. Description of the invention (91

Q—X

And the diketone product conforms to formula XIA

XIIA

Γ 2 I c = x χΙΑ where -A-A-, -B-B, Υ1, Υ2, and X are as defined in Formula VIIIA, respectively. Further according to plan 2, the dione of formula XI is reacted with an alkali metal alkoxide to form Mexileau or other products conforming to formula X

Printed by the Staff Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs

X wherein -A-A-, R3, -B-B ·, R8 and R9 are as defined in formula VIII, respectively. R1 is as defined in Formula V. This method uses roughly the same formula as above. -94- This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) 593305 A7 5

, Description of the invention (92)

The conversion of the V1 compound to those of the formula V is accomplished by the same reaction plan. Preferably, the enzyme of formula XI conforms to formula XIA

-c—X ΧΙΑ and the intermediate product conforms to the formula ⅩΑ (Please read the precautions on the back before filling this page)

CCH2D1 -c = x ΧΑ Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economics where-Α_Α_, _Β_Β_, γΐ, γ2, and χ are as defined in the formula XIII. R1 is as defined in Formula V. Next, by a novel biotransformation method, Conreno and other compounds of formula χ are hydroxylated to produce the product of formula IX. -95- This paper is suitable for paper size ® CNS A4 (210X; 97mm) 593305 A7 B7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs. 5. Description of the Invention (93)

Wherein-AA-, R3, -BB-, R8 and R9 are as defined in Formula VIII, and R1 is as defined in Formula V. The organism that can be used in this hydroxylation step is soil mycelia (Nocardia conicruria) ATCC 3 1548, Nocardia aurentia ATCC 12674, Corynespora cassiicola ATCC 16718, Streptomyces hydroscopicus ATCC 27438, Mortierella isablellina ATCC 42613 &gt; Beauvria bassiana ATCC 7519, Penicillum purpurogenum ATCC 46581, Hypomyces chrysospermus IMI 109891, Tamnostylum piriforme ATCC 9899, short spines Cunnignhamella blakesleeana ATCC 8688a, Cunningnhamella echinulate ATCC 3655, Cunninghamella elegans ATCC 9245, Trichothecium roseum ATCC 12543, Hermicilla Epicoccum humicola ATCC 12722, Saccharopolyspora eythr ae) ATCC 1 1635, Beauvria bassiana ATCC 13144, Arthrobacter simplex, Bacterium cyclooxydans ATCC 12673, Radisi-96- This paper is in accordance with China National Standard (CNS) A4 Specifications (21 0X 297 mm) (Please read the notes on the back before filling out this page),? Τ 593305 A7 B7 V. Description of the invention (94) Cylindrocarpon radicicola ATCC 11011, Orange soil mycelia (Nocardia aurentia) ATCC 12674, Nocardia canicruria, Norcardia restrictus ATCC 14887, pseudomonas testosteroni ATCC 1 1996, horse Rhodococcus equi ATCC 21329, Mycobacterium fortuitum ATCC-6842 and Rhodococcus rhodochrous ATCC 19150. This reaction is generally carried out by the method described above with regard to Figs. 1 and 2, which is particularly preferred. The growth medium used in the biotransformation preferably contains available nitrogen between about 0.5% to about 5% by weight; glucose between about 0.5% to about 5% by weight ; Yeast derivatives between about 0.25% to about 2.5% by weight; and available phosphorus between about 0.05% to about 0.5% by weight. The best growth media include the following: Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling out this page) Soy meal: between about 0.5% to about 3% by weight Glucose; soy meal between about 0.1% and about 1% by weight; metal halide between about 0.5% and about 0.5% by weight; between about 0.5% and 0.5% by weight Yeast derivatives between about 56.5% by weight, like autolyzed yeast or yeast extract; phosphates, between about 0.05% by weight and about 0.5% by weight, like K2HP〇4 pH = 7; glandular-yeast extract-glucose: glands between about 0.2% and about 2% by weight; yeast extraction between about 0.5% and about 0.5% by weight And glucose between about 2% by weight and about 5% by weight; -97- This paper size applies to China National Standard (CNS) A4 (210X297 mm) &quot; 593305 A7 B7 V. Description of the invention (95 Leellerton (Mueller-Hinton); at about 10% by weight Bovine infusion between about 40% by weight; casein amino acid between about 0.35% by weight and about 8.75% by weight ° / 〇; between about 0.5% by weight and about 0.7% by weight It can be grown in soybean meal or glandular nutrients, while Actinomyces and tadpole bacteria can be grown in soybean meal (for biotransformation, add 0.5% to 1% by weight Carboxylate, such as sodium formate) or Geller_Hydron broth. J j Lu-Hydroxy mexanol produced from Mex Renault by fermentation is discussed in Example 19. Product of Formula IX Are novel compounds that can be isolated by filtration, washed with a suitable organic solvent, such as ethyl acetate, and recrystallized from the same or similar solvents. They have the ability to prepare compounds of formula I, especially formula IA The actual valence of the intermediate of the compound is 较佳. Preferred compounds of formula X are in which -Α_Α · and -BB- are _CH2_CH2 ·, R3 is hydrogen, a low-carbon alkyl group or a low-carbon alkoxy group, and R8 and R9 —To form a 20-spiroxane ring:

XXXIII (Please read the notes on the back before filling out this page} Order In the next step of Synthesis Plan 2, the product of formula IX is reacted with a dehydrogenation reagent to produce a compound of formula 11

II -98- This paper size is in accordance with Chinese National Standard (CNS) A4 specification (21〇X 297mm) 5. Description of the invention (96 A7 B7 of which -AA-, R3, β R 1, n /--V λ / … R and Han 9 are as defined in Formula VIII, and R is as 佘 # in Formula V, meaning ^ When the subject enzyme conforms to Formula IXA,

The product is of formula IIA

IXA C # first read and read the back, the notes really fill in this page)

C〇H2 ^ -c-x

IIA The Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs printed -AA-, -BB-, V1, and R1 is defined as 1 in formula V and X is defined as in formula XUI, respectively. The product such as the compound of formula I is converted by epoxidation by the method described in US Patent No. 4,559,332; or preferably by the novel epoxidation method of the present invention described in the foregoing. In a particularly preferred embodiment, the complete method of the Plan 2 process is as follows: -99- This paper size is suitable for financial and related standards (CNS) &amp; See the grid (210X297 public ¢ 7 593305 A7 B7 V. Description of the invention (97

Conreno

Printed by the Staff Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs

(Please read the precautions on the back before filling this page) -100- This paper size is applicable to the Chinese National Standard (CNS) A4 specification (210X 297 mm) 593305 Α7 Β7 V. Description of the invention (98) jtjj 3 In this case Synthesis begins with the enzyme

Where-Α-Α- and R3 are the same as defined in formula vm, and β_β_μ is the same as in formula VIII, except that R6 and R7 are not in μ, and part of the ring fused with ring 0 at position 7 And R26 is a low carbon number alkyl group, preferably a methyl group. Reaction of the parent enzyme of formula XX with sulfonium ylide to produce an epoxy intermediate conforming to formula XIX

(Please read the precautions on the back before filling out this page), 11 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs XIX Among them, R3, _B_B_ * R26 are as defined in the formula χχ. In the next step of Synthesis Plan 3, the intermediate of formula χιχ is transformed into another intermediate of formula XVIII-101- This paper size applies the Chinese National Standard (CNS) A4 specification (210 × 297 male thin) 593305 Central Ministry of Economic Affairs Printed by the Bureau of Standards Consumer Cooperatives V. Invention Description (99 A7 B7

CO.Et XV workers where -A-A-, R3 and 4 n 4 — B- are as defined in formula xx. In this step, spermidine reacts with NaCH (CoPt, 丄,), 2 in the presence of a base in a solvent, and converts the enzyme of formula χΐχ into an intermediate of formula xvm. A compound of formula χν⑴ + in ’,’, water, and a metal-detecting compound, yields an unbroken intermediate compound conforming to formula mi Ο

XV 工 工 where-Α · Α ·, R3 and as defined in the formula χχ. The method of converting a compound of formula χχ to a compound of formula XVII is basically equivalent to those described in U.S. Patent Nos. 3,897,417, 3,413,288, and 3,330,489, which are specifically incorporated herein by reference. Although the enzymes are different, the reagents, mechanisms and conditions for introducing 17-spirolactone are basically the same. The reaction between the intermediate of formula XVII and the dehydrogenation reagent produces a further intermediate of formula XVI. 102- This paper size applies the Chinese National Standard (CNS) A4 specification (2) 0 × 297 mm. (Please read the note on the back first (Fill in this page again)

1T 593305 A7 B7 V. Description of the invention (100) 〇

XVI where-A-A-, R3 and -B-B- are as defined above. Representative useful dehydrogenation reagents include dichlorodicyanobenzoquinone (DDq) and chloroquine (2,3,5,6-tetrachloro-p-benzoquinone). In addition, the dehydrogenation can also be accomplished by the _ation at carbon_6, followed by the dehydrogenation. Next, the intermediate of formula XVI is converted to amidine of formula XV. Please read the precautions on the back before filling in this page.

νηλ-Order printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs

XV in which Α-Α-, R3, and Β_Β are as defined in Formula XX. The conversion is basically a method for converting the 11α-hydroxy compound of the formula VIII to the amidine of the formula νπ by cyanation. Usually the source of cyanide ions can be metal cyanide. The base is preferably pyridine and / or tetramethylguanidine. Methanol solvents can be used. -103- I Paper Size Appropriate (CNS) A4 Specification (21〇, χ 297 mm D &quot; ~ 593305 A7 B7 V. Description of the Invention (101) The product of formula X v is a newly reduced compound 'can be obtained by layer These and other novel compounds of formula AXV have practical valence as intermediates for the preparation of compounds of formula I, especially of formula IA. Compounds of formula AXV conform to the following structure

B k

AXV where-A- A-, -B-B-, R3, R8 and R9 are as defined above. In the most preferred compound of formula X V, '-and-are -CH2-CH2-. According to the hydrolysis of the diketone compound of formula VI above, the enamine of formula χν can be converted to the diketone of formula XIV. (Please read the precautions on the back before filling this page)

、 1T Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 〇

XIV where -A-A-, R3 and -B-B- are as defined in formula XX. About Epoxy -104- This paper size applies to Chinese National Standard (CNS) A4 specification (21〇 × 297mm) 593305 A7 B7

V. Description of the invention (102 For the synthesis of Max Reynolds, the compounds of formula XIV that also fall within the scope of formula VIA are particularly preferred. The products of formula XIV are novel compounds that can be separated by precipitation. These and Other novel compounds of the formula AXIV have practical valences as intermediates for the preparation of compounds of the formula I, especially of the compounds of the formula IA. The compounds of the formula AXIV conform to the following structure

AXIV where-A_A_, _B_B_, R3, R8 * R9 are as defined above. In the most preferred compounds of the formulae AXIV and XIV, · A-A &gt; _Β-Β- is -CH2_CH2-. The compound of formula XIV is further converted to a compound of formula XXXI basically using the method of converting a diketone of formula VI to a hydroxy ester of formula V described above. In this case, ‘must be converted to the product of formula χχχΠ (please read the notes on the back before filling this page)

Printed by the Staff Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs

First isolate the intermediate formula XXXI

XXXI -105- This paper size applies to Chinese National Standard (CNS) Α4 specification (210 '乂 297 mm)

Heating 593305 Α7 Β7 V. Description of the invention (103) XXX workers-where A-A · and _B-B-are as defined in formula XX. Preferred compounds of formula XXXI are those within the scope of formula IIA. The compound of formula XXXI is converted to a compound of formula XXXII using the method described above or the method in U.S. Patent No. 4,559,332. In a particularly good embodiment, the complete method of the plan 3 process is as follows: (Please read the precautions on the back before filling this page)

Printed by the Staff Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs

HCI, CH # OH, ΗλΟ

This paper size applies the Chinese National Standard (CNS) A4 specification (210 × 297 mm). 5. Description of the invention (104 A7 B7 itill gate 1ΓΓ 骤 and ㈣3㈣, which is the formula of the compound χχ starting from the meaning of the compound. The intermediate of xvn. 1¾►Then φρη% of the formula X V11 Luta a 4 559 Chuan ... For example, using the method of National Patent No. 9,332, to produce a compound of formula XXIV.

(Please read the notes on the back before filling out this page). Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs XXIV 2_a-a-, R ^ _B_B • As defined in the formula xtt. However, in a particularly preferred embodiment of the invention, according to the method described in the above paragraph, the method of converting the formula to a product of 3U, using an oxidant including a lye peroxide activator, the most preferred is trichloroethyl Ammonium amine, spanning 9, &quot; · Double bond epoxidizes the formula χνπ by the enzyme. The conditions and reagent ratios of this reaction are roughly the same as those described in the formula II # g to epoxy Mexileau. It has been found that peroxyacids, such as, for example, m-chloroperoxybenzoic acid, can be used to complete the enzymatic epoxidation of formula XVII in good yields. However, trigas acetamidine reagents provide excellent results in reducing the formation of Bayer-Villager oxidation by-products. The latter by-product can be removed, but this requires trituration from a solvent such as ethyl acetate, which is then crystallized from other solvents such as dichloromethane. Dehydrogenate the oxygenated compound of formula XXIV by reacting with a dehydrogenating (oxidizing) agent such as DDQ or chloroquinone, or by using bromination / dehydrobromide (or -107- this paper standard applies) China National Standard (CNS) Α4 specification (210X297 mm)

1T I -1. 593305 A7 B7 V. Description of the invention (105) The procedure of other halogenation / dehydrohalogenation) produces a double bond between 6- and 7-carbons, and produces other compounds of formula XXIII. Novel intermediate

XXIII Printed by the Consumer Standards Cooperative Bureau of the Central Bureau of Standards of the Ministry of Economic Affairs where -A-A- and -B-B- are as defined in formula XX. Particularly preferred compounds of formula XXIII are those in which -A-A · and -B-B are as defined in formula XIII. Although direct oxidation is effective for forming the product of formula XXIII, the yield is usually low. Therefore, it is best to complete the oxidation in two steps. First, the formula XXIV is halogenated by the enzymatic substance at the C-6 position, and then dehydrogenated to 6,7-chain pyrene. The halogenation is preferably carried out using an n-halogen organic reagent such as, for example, N-bromosuccinimide. In a suitable solvent, such as, for example, acetonitrile, it is brominated in the presence of a initiator of a chemical reaction such as benzamidine peroxide. The reaction can be efficiently performed in a temperature range of about 50 to about 100 ° C, conveniently at a reflux temperature of atmospheric pressure, in a solvent such as tetragasified carbon, acetonitrile, or a mixture thereof. However, in order to complete the reaction, it is usually necessary to carry out the reaction for 4 to 10 hours. The reaction solvent is stripped off and the residue is dissolved in a water-immiscible solvent such as ethyl acetate. The resulting solution is continuously rinsed with a mild alkaline solution (such as an alkali metal bicarbonate) and water, or preferably saturated brine, to reduce product loss. The solvent is then stripped and the residue is dissolved in other suitable solvents. (Please read the notes on the back before filling this page), 11 -108- Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 593305 A7 _____ B7 V. Description of the invention (106) Solvents for dehydrogenation reactions (like Dimethylformamide). Add appropriate dehydrohalogenating reagents, such as 丨, 4-diazabicyclo [2,2,2] octane (DABCO), alone or with an alkali metal halide such as LiBr Medium 'heats the remote solution to an appropriate reaction temperature, such as 60 ° C to 80 ° C, and continues the reaction for several hours, usually 4 to 5 hours, in order to effect dehydrobromide. If necessary, additional dehydrobromide reagents can be added to the reaction cycle to complete the reaction. The product can then be precipitated by, for example, adding water, and then separated by filtration. The product of formula XXIII is recovered, preferably washed with additional water. Preferably, the product is recrystallized from, for example, dimethylformamide. Products of formula XXIII, such as 9,1 1-epoxy Conreno, are novel compounds that can be separated by extraction / crystallization. They have practical valences as intermediates for the preparation of compounds of formula j, especially compounds of formula IA. For example, they can be used as enzyme substrates for preparing compounds of formula X X11. In the most preferred compound of formula XXIII, -A-A- and -B · B · are -CH2-CH2-. Using the above method for preparing a compound of formula VII on large fa, the compound of formula X X111 is reacted with a cyanide ion to produce a novel epoxyenamine compound in accordance with formula XXIIi

This paper size applies to China National Standard (CNS) A4 (210X297 mm) (Please read the precautions on the back before filling this page)

593305 A7 V. Description of the invention (107) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs where-A-A-, R3 and · Β-Β-as defined in Formula XX. Particularly preferred compounds of formula XXII are those in which -A-A- and -B-B- are as defined in formula XIII. The product of formula XXII is a novel compound which can be separated by precipitation and filtration. They have practical valences as intermediates for the preparation of compounds of formula I, especially compounds of formula IA. In the most preferred compound of formula XXII, -A-A- and -B-B- are -CH2-CH2-. In general, the method for preparing the compound of formula VI is used to convert the epoxy enamine compound of formula X X11 into a novel epoxy diketone compound of formula XXI. The product of formula XXI is a novel compound which can be separated by precipitation and filtration. They have practical valences as intermediates for the preparation of compounds of formula 丨 and especially compounds of formula IA. Particularly preferred compounds of formula XXI are those in which eight and eight are as defined in formula XIII. Among the best compounds of formula χχι, -Α-Α- and -B-B ^ _CH2_eH2_. Using the epoxidation described above, or a compound of formula XX! Into a compound of formula in US Patent No. 4,559,332. In the specific embodiment of the best, the complete method of the plan 4 process is as follows: -110- This paper size is applicable to Chinese national standards ((Please read the precautions on the back before filling this page) 丨 «. Order-^ 1. 593305 A7 B7 V. Description of the invention (108)

(Please read the notes on the back before filling this page)

Printed by 1T Consumer Cooperatives, Central Standards Bureau, Ministry of Economic Affairs

-111- This paper size is in accordance with Chinese National Standard (CNS) A4 specification (210 × 297 mm) 593305 A7 B7 V. Description of the invention (109 plan Plan 5 The method of the plan starts with the enzymes conforming to the formula X XI X Λ 〇

XXIX where _Α_Α_ and · Β-Β_ are as defined in Formula XX. The enzyme is converted into a product of formula XXVIII by reaction with trimethyl orthoformate.

XXVIII. Where-Α-Α-, R3 and -Β-Β- are as defined in formula XX. After the formula XXVIII is formed, a compound of the formula XXIX is converted to a compound of the formula XXVII by the above-mentioned method for converting an enzymatic substance of the formula XX to a formula χνιι. The compound of formula XXVII has the following structure: (Please read the notes on the back before filling out this page. J Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs.

XXVII where -A-A- and -B-B- are as defined in formula XX, and Rx is any common hydroxy protecting group. This paper size applies to China National Standard (CNS) A4 (210X297 mm)

-112- 593305 A7 B7

V. Description of the invention

XXVI where -A-A- and -B-B- are as defined in formula XX. Particularly preferred compounds of the formula χχιχ, XXVIII, χχνπ and XXVI are those in which _A_A- and _b_b_ are as defined in formula XIII. The product of formula XXVI is a novel compound which can be separated by precipitation / filtration. They have practical valences as intermediates for the preparation of compounds of formula 尤其, especially compounds of formula AA. Particularly preferred compounds of formula XXVI are those in which _A_A_ and -B-B- are as defined in formula XIII. In the best compound of formula χχνι, -A_A- and. Using the above method for cyanation of compounds of formula VIII, the novel intermediate of formula XXVI is transformed into a novel intermediate of hydroxyxamine of formula χχν (please read the precautions on the back before filling this page), 11 Ministry of Economy Printed by the Central Bureau of Standards Consumer Cooperatives

XXV -113- This paper size applies Chinese National Standards (CNS) Rule 8 ^ 771 ^ 297 305 593305 Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs Α7 Β7 V. Invention Note (ill) Among them -AA-, R3 * _B_B_ is as defined in Equation XX. The product of formula XXV is a novel compound which can be separated by precipitation / filtration. They have practical valences as intermediates for the preparation of compounds of formula I, especially compounds of formula IA. Particularly preferred compounds of formula XXV are those in which _A-A_ and B- are as defined in formula XIII. Among the best compounds of formula χχν, -A-A- and -B-B- are -CH2-CH2-. Basically the conditions described above for the preparation of diketone compounds of formula VI were used to convert the 9-hydroxyenamine intermediate of formula XXV to the diketone compound of formula XIV. In this case, it is noted that this reaction can efficiently hydrolyze the enamine structure simultaneously and dehydrate at the position of 9, 1 1 in order to introduce a 9, 丨 丨 double bond. The compound of formula XI v is then converted to a compound of formula XXXI using the same procedure as in Scheme 3 above, and from it to a compound of formula χIII. In a particularly preferred embodiment, the complete method of the Plan 5 process is as follows:

Epoxy Mex Renault-114- This paper size applies to Chinese National Standard (CNS) A4 (210X 297 mm) 593305 A7 B7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the Invention (112) Plan 6 Ten d 6 kt, an advantageous method for the preparation of epoxy mexanolol and other compounds according to formula I, starts with 1 1 α-hydroxylation of androstenedione or other compounds according to formula χχχν

XXXV where · Α · Α-, R3, and -Β-Β_, as defined above, produce intermediates conforming to j X X X VI

XXXVI where _A_A_, r3 * _B-B- are as defined in the formula χΠΙ *. Except for the choice of enzymes, the method of operating 1 1 α-hydroxylation is basically the same as described in Plan 1 above. The following microorganisms can perform 1 1 α-hydroxylation of androstenedione or other compounds of the formula XXXV: Aspergillus ochraceus NRRL 405 (ATCC 18500); Aspergillus tiiger, ATCC 1 1394 ; AspergHlus nidulans ATCC 1 1267; -115 This paper size applies to China National Standard (CNS) A4 specification (210X297 mm) {Please read the precautions on the back before filling this hundred

A7 ____ B7 V. Description of the invention (ll3) 2S-M bacteria (Rhizopus orvzae, ATCC 11145; Rhizopus stolonifer, ATCC 6227b;

Spore end (Ijichothecium roseurrn ATCC 12519 and ATCC 8685). Next, by using 1 1 a -hydroxyandrosta-4 -fluorene-3,17 -dione or other compounds of formula XXXVI, such as trialkyl orthoformate Etherification reagents such as these are reacted in the presence of an acid catalyst to transform them into 丨 丨 "of formula (丨 〇 丨) _ hydroxy-3,4-fluorenol ether: 3

(Please read the notes on the back before filling this page) 101 where -AA-, -R3, and -Β-Β- are the same as defined in formula XIII, and R11 is a methyl group printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs Or other low carbon number alkyl (Ci to C4). To complete the conversion, acidify the 1 1 α-hydroxyl enzyme by mixing it with an acid, such as benzene ~ hydrate or toluic acid hydrate, and dissolve it in a low-carbon alcohol solvent. Good is ethanol. Over a period of 5 to 40 minutes, the tri- orthoformate, preferably triethyl orthoformate, is gradually introduced, while maintaining the mixture in the cold ', preferably at about 15 to about 15x :. The mixture was then allowed to warm and the reaction was completed at a temperature between 20 ° C and about 60 ° C. It is best to complete the reaction at 30 to 50 ° C for 1 to 3 hours and then heat to reflux for an additional time, usually 2 to 6 hours, to complete the reaction. The reaction mixture is cooled, preferably 0 to 15 ° C, and most preferably about 5 ° C, and the solvent is removed at 眞 S. -116- This paper size is applicable to China ^^ (CNS) A4 specification (210X297 mm)-"W3305 Printed by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs, and the description of the invention (114) Use the same as above in Plan 3 The reaction plan is to convert a compound of formula XX into a compound of formula XVII, and introduce a part of 17-spirolactone of formula χχχπι into the compound of formula 101. For example, formula 101 can be reacted with an enantiomeric salt in the presence of a base such as metal chloride, in a suitable solvent such as DM s0, to produce an intermediate conforming to formula i 0 2 Things:

Where _A-A-, R3, Rl1, and are as defined in Formula 101. Then, the intermediate of Formula 102 &lt; and the malonic diester are reacted in the presence of an alkali metal alkoxide to form a five-membered spironolactone ring, and an intermediate of Formula 103 is produced

Where -n, R3, RU, R12, and _b_b- are as defined in the formula χιπ. Finally, the compound of formula 103 is subjected to heating in an appropriate solvent, such as dimethylformamide, in the presence of an alkali metal dentate, so that the alkoxycarbonyl group is partially cleaved, to give formula 104 Intermediate · -117- This paper size applies Chinese National Standard (CNS) A4 specification (77 ^^^) (Please read the precautions on the back before filling this page)

593305 A7 B7 V. Description of the Invention (Export 〇

104 where -A-A-, R3, R &quot; and -B-B · are again as defined in the formula χπι. Next, the 3,4-fluorenol ether compound! 〇4 is transformed into a compound of the formula χχπι, that is, a compound in which R8 and R9 together form a part of the formula χχχιπ. This oxidation step is basically performed in the same way as in the synthesis of Scheme 4, the oxidation of a compound of formula XXIV into an intermediate of formula XXIII. Direct oxidation can be accomplished using reagents such as 2,3_digas_5,6_dicyano-, 4-benzoquinone (DDQ) or tetrachlorobenzoquinone (chloroquinone), or better It is brominated by first using a bromine agent such as N • bromosuccinimide or N-, 3-dibromo-5,5-dimethylendourea (DBDMH), and then using The base will debromize it, such as sLiBr and heat in the presence of DABCO, 'to complete the two-step oxidation. Where nb s is used for bromination, acid must also be used to convert 3-enol ether to ketene \ DBDMH, an ionic rather than free radical bromination agent, which is effective for bromination in itself, And convert the methyl alcohol ether to ketene. Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs: ft Use the ^ described in Plan 1 above to convert ^ VI11 compounds to epoxy mexrenol, or other compounds of formula. The intermediates of formulae 1, 1, 102, 103, and 104 are all novel compounds with actual valences as epoxy intermediates of Mexrenault or other compounds of formulae IA and I. In each of the compounds of formula 101, 102, 103, and 104, -AA- and _Β · Β_ are preferably -cuHr, and R3 is hydrogen. (CNS) M specification (2) () &lt; 297 public attack -118-593305 A7 B7 5. Description of the invention (116) Alkyl or low carbon number alkoxy group, preferably the compound of formula 101 is 3 -Ethoxy-1 1 α -hydroxyandrosta-3,5-diene-1 7-one, the compound of formula 102 is 3 _ethoxyspiro [androsta-3,5-diene-1 7 / ?, 2 '-ethylene oxide] _ 1 1 π -alcohol, the compound of formula 103 is 3 -ethoxy-11 α, 17α -dihydroprogestin-3,5_diene_ 21,21- Ethyl hydrogen dicarboxylate, r-lactone, and the compound of formula 104 is 3-ethylethoxy-11 ubiquitin, 17 α --- Mouse®-3, 5--Women-2 1-: ¾ 'τ &quot;-内 酉 定 0 In a particularly preferred embodiment, the complete method of the plan 6 process is as follows:

(Androstenedione CAP)

Heating (Please read the notes on the back before filling out this page)

N «CI, DMF

-119- This paper size is in accordance with Chinese National Standard (CNS) A4 (210X 297 mm) 593305 A7 B7 V. Description of the invention (in) _ 丨 丨 ___j ----- Program 7 Program 7 provides epoxy beauty Synthol and other compounds of the formula are synthesized using starting enzymes including θ-glutanol, cholesterol, stigmasterol or other compounds of the formula

R1 Printed by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. rh, R15, and R16 are each selected from hydrogen or Ci to c4 alkyl. In the first step of the synthesis, 1 1 α-hydroxyandrostenedione or other compounds of the formula χχχν is prepared by biotransformation of the formula χχχνπ. In general, the biotransformation process is completed according to the above-mentioned method of 11_ hydroxylation of Conreno (or other enzymes of formula XIII). In the synthesis of 1 1 α-hydroxyandrostenedione, 4_androstil 7 • dione was initially prepared by the biological transformation of a compound of the formula χχχνπ. The initial biotransformation 'can be accomplished by the method described in U.S. Patent No. 3,759,791, which is specifically incorporated herein by reference. Subsequently, according to the above-mentioned method of α-hydroxylation of Conreno (or other enzymatic substance of formula XIII), 4-androstidine _ 3, i 7 _ dione was converted to 丨 α _ Hydroxyandrogens are associated with dioxins. The rest of Program 7 is the same as Program 6. In the special embodiment (please read the notes on the back before filling this page)

-120- 593305 A7 B7 V. Description of the Invention (118) The complete method of the plan 7 process is as follows:

(Please read the precautions on the back before filling out this page) The major complication of the synthesis of Epileur Reynolds and related compounds printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs requires the stereoselective introduction of α at 7-carbon -Alkoxycarbonyl substituents without unwanted modification elsewhere in the steroid structure. According to the present invention, an effective synthetic route for the introduction of a 7 α-alkoxycarbonyl group has been disclosed, which involves cyanation at 7-carbon at first, followed by the formation of 5,7-dilactone, and the dilactone and alkylating agent The base opens the reaction to give the desired 7α-alkoxycarbonyl substituent. -121-This paper size is in accordance with Chinese National Standard (CNS) A4 specification (210 × 297 mm) 593305 A7 B7 5. Description of the invention (l20) 7 acid is removed in its salt form. In order to form dilactone, known methods can be used to prepare the 7-cyanoenzyme. The unsubstituted enzyme at the 7-carbon and a slight excess of chaotic ion can be used, preferably per equivalent. The substrate is reacted in a weakly acidic solution containing a water / DMSO solvent mixture, with a large W.G5 to about κ25 equivalents. Preferably, the reaction mixture contains a carboxylic acid, such as about one equivalent of acetic acid per equivalent of the enzyme. 7a_ * 70_CN isomers are formed, with 7 pan-isomers as the main isomers. 7-cyanosteroids can be recovered using conventional methods. The accessory can be prepared using other methods known in the art. Usually according to plan 8, can be substituted with copper, R8 or R9 at the 17-position, where R8 and R9 are as described above, and have aliphatic, alkenyl, epoxy or hydroxyl substituted configurations at C-9 and C-11 Type 7-cyano intermediates to form 5,7-dilactones, that is, (please read the precautions on the back before filling in this page)

XL one &gt;

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% 90 This paper size applies to Chinese National Standard (CNS) A4 specification (210X297 mm) 593305 Μ B7 V. Description of the invention (121) Among which -AA-, -BB- and R3 are as defined above, R80 and R90 and R8 Same as R9, or R8G and R9G together form a ketone, and R18 is as described below for plan 9, and _ E-E-is selected from

For example For example VCH- CH, XL workers &quot; C-CH- OH 'CH- CH-

XLIV

XLV (Please read the notes on the back before filling this page) «·

, 1T on24 \ I c—- For example; c—ch2.

XLVI 〇 ”一 · Usually &gt; 1η ·

XLVII then converts the compound of formula XLII to 7 a · alkoxy_carbonyl: printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

XL · Gonggong -124- This paper size applies to Chinese National Standard (CNS) A4 (210X 297 mm) 593305 A7 B7 V. Description of the invention (I22) R β · 〇

XLVI workers In each of XL, XLI, XLII and XLVIII, R80 and R90 preferably form a ketone or / / ί 7) IIIMI (CHg) 4

XXXIV where Υ1, Υ2, X and C (17) are as defined above, preferably

(Read the notes on the back before filling in this page)

Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs of the 1T. XXXIII R3 is preferably Η, R1 is preferably methoxycarbonyl, and -A-A- and -B-B_ are each preferably -CH2-CH2-. It will be understood that the reaction can also be accomplished using a protected 3-keto group by converting it and maintaining the form of each ether or ketal throughout the reaction sequence. Another method of plan 8 involves, as mentioned earlier, the use of various intermediates in the range of formulas XLI and XLII. It is noted that via Scheme 8, the reagent for forming 5,7-dilactone from 3-keto-A · 4,7-carboxyl group is trisorcinyl orthoformate. In Plan 6, the 1 1 α-transylandrostidine The conversion of the diketone to 3-alcohol ether-Δ-1 1々 · hydroxy intermediate i 1 also uses the same reagents. It is believed that the route of the reaction of plan 8 depends on the substitution at c-7. Reacts with orthoformate in the presence of H + to form an intermediate carbohydrate ion. 125 This paper is sized to the Chinese National Standard (CNS) A4 (210X297 mm) 593305 A7 _________ B7 5. Description of the invention (123) It has a carboxyl group at C-7, and its positive charge is balanced between c_3 and c_5. When the proton is lost, the C-3 carbohydrate ion produces a compound of formula 101, and the c-5 carbohydrate ion generates a dilactone. There is hydrogen at C-7, and it is believed that 3,5-diene-3-alkoxy (enol ether) is preferred because of its double bond conjugation. Carbion ions with a 7a-C02 substituent c_5 at cj are captured by the carboxyl group and formed into lactone. At this time, the 3-keto I group is preferentially converted to a ketal, thereby driving the reaction to completion. In the following plans 9 and 10, preferred specific embodiments of plan 8 are described. Plan 9 4 Plan 9 begins with the same enzymes as Plan 4, namely compounds of formula X X. First oxidize the enzyme to a compound of formula B: (Please read the precautions on the back before filling this page) R3 η

The Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs, Yin Li, where -A_A-, R3 and -B-B- are as defined in Formula XIII. The oxidation reaction is operated according to any of the above reaction schemes related to the conversion of a compound of formula χχι v to an intermediate of formula χχιπ in the synthesis of Scheme 4. Using the method described in Scheme 8, convert the compound of formula B to the 7-cyano intermediate of formula c: R3

The scale of this paper applies the Chinese National Standard (CNS) A4 specification (210 × 297 mm) 593305 A7-· -____. B7 ^ 一 &quot; 一 —— 一 _ 5. Description of the invention (124) of which Α-Α-, &amp; 3 and _BB- as defined in Formula XIII. The compound of formula c is then converted to a 5,7-lactone of formula D using the trialkyl orthoformate reagent previously used in Scheme 6: R3

(Please read the notes on the back before filling in this page. _A_A_, R3, and _B_B_ are defined in sXIn *, and r17 is C 1-C 4 yuan basis. ★★ Hei-style D of 5,7 _ two internal @purpose, for example, by removing the acid by bicarbonate washing, it can be easily separated from the unreacted 7-cold-c 〇η ′, thereby establishing Desired C-7 stereochemistry and hinder epimerization in subsequent reactions operating under alkaline conditions. As described in Plan 8, the lactone is reacted via reaction with an alkyl group Esterification yields a vinegar intermediate of formula j. Continue the synthesis of plan 9 to convert a compound of formula D to a compound of formula 11. The protector with a 3-keto group has been converted to a ketal, according to plan 3 and above The reaction plan of 6 is that the 20-spiroxyalkyl group of formula XXXIII is selectively introduced to the 1 7-position, thereby generating a compound of formula E,

This paper size is in accordance with Chinese National Standard (CNS) A4 specification (21 × 297 mm), but can be printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs, A7, B7. 5. Description of the invention (125) Because 3-ketones are protected, they can Choose hydrolysis conditions that are best suited to attack C-1 7 _ and have nothing to do with the formation of by-products by reaction at the 3-position. After the 3-ketal compound of formula E is hydrolyzed to a 3-keto group structure of formula F,

The latter intermediate is reacted with an alkyl iodide in the presence of a base, via the conversion of Scheme 8, to produce an intermediate ene ester of formula II. Finally, using any of the methods in Scheme 1 above, the latter intermediate is converted to epoxy mexrenol or other compounds of formula I. Plan 9 not only benefits from controlling the stereochemistry supplied by the 5,7-lactone intermediate, but also gains the advantage of allowing a wide range of hydrolysis conditions without hindering 17-spirolactone. Like the reactions of the other synthetic schemes of the present invention, the reactions of Scheme 9 can also be used; different enzyme-converting effects than those specifically described above. Thus, for example, a compound substituted at the 17-carbon with Han 8 and R9 as defined above, or more specifically a substituent of the formula:

Eight I

C (1 7) IIUK CH2) 2. Q-X ^, 'χχχιχ /, medium, γ1 and Y2 are as defined above, and c (17) represents 17_ carbon, on the compound substituted at 17_ carbon, Complete 3_keto · or 3_ketal cyano-solids_ -128- This paper is suitable for financial standards (CNS) M specifications (2lQxw)

593305 A7 B7, description of the invention (I26 alcohol to 3-keto- or 3-ketal_5,7_ dilactone conversion, or 3 · keto- or 3-ketal, 7-dilactone to 7 Conversion from -alkoxycarbonyl. However, by using a 17-ketone-specific enzymatic reaction procedure, and the subsequent introduction of the above-mentioned 17-spirolactone and 7-s-oxyalkoxy groups into 3_keto- △ _ 9, 1 1 specific response plan for steroids, and gain significant benefits, especially in the economics of the process. Γ Formulas D, E, and F are novel compounds that are synthesized according to plan 9, which It can be used to prepare epoxy mexanol and other compounds of formula I and IA. Among these compounds, -AA- and _B-B_ are preferably · 〇Η2 · 〇Η2_, and r3 is hydrogen and low carbon number Alkyl or low-carbon alkoxy. The best compounds of formula D are those in which R17 is methoxy. Eight In a particularly preferred embodiment, the complete method for the process of plan 9 is as follows. (Please read the note on the back first Please fill in this page again) Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs -129- This paper size is applicable to China National Standard (CNS) A4 (210X297 mm) 593 305 B7 V. Description of the invention (127) Ο Ο

(Please read the notes on the back before filling out this page) Order Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs -130- This paper size applies to China National Standard (CNS) Α4 size (210 × 297 mm) 593305

DESCRIPTION OF THE INVENTION (l28 it, Division 1 ο 7 to cyano intermediates in Form C to Form 10 are the same as in Plan 9. In the next step of Plan 10, 7-cyanosteroids and trialkyl orthoformate The reaction is carried out in an alkanol solvent, preferably in methanol with trimethyl orthoformate, while protecting these two groups by converting 3-ketone to methyl alcohol and 17-ketone to ketal. Subsequently, for example, by reacting with a dialkylaluminum hydride, preferably diisobutylaluminum hydride, the 7-cyano group is reduced to a 7-formamidine group, thereby producing a compound of formula 203: 203 of which-AA -, R3 and _B_B_ are as defined in the formula χΠI, and Ri8 is CpC: 4 alkyl. Before protecting the keto group, as described above, its reduction is prevented by hydrogenated dialkyl group. Then formula 2 is used. The intermediate of 3, in the presence of excess alcohol (R19OH), reacts with the diluted aqueous acid to selectively hydrolyze the 17 ketal to produce the intermediate of formula 2.04: (Please read the precautions on the back first (Fill in this page again) Order O'9 printed by the Consumers' Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs

[204] where R19 is selected from low carbon number alkyl (preferably CiSC4), or R19 group is in -131- This paper size is applicable to Chinese National Standard (CNS) A4 specification (210x 297 mm) 593305

Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs ^ 〇

CO ^ R A7 B7 V. Description of the invention (Π9) A cyclic 0,0-oxoalkoxy substituent at the 3-carbon position at the 3-position. Furthermore, the hemiketal was protected by treatment with alkanol (R19〇H) in the presence of a non-aqueous acid [204] to produce an intermediate of formula 205: [205] where-A- A_, -BB -, R3 and R19 are as defined above, and R20 is a C to C 4 alkyl group. Then according to the above reaction steps of plans 3 and 6, 1 7 _ spironolactone is introduced, so proceed through the procedure outlined below:

2S

[207] 4 -132- This paper size applies to Chinese National Standard (CNS) A4 (210X297mm) (Please read the precautions on the back before filling in this page}

593305 A7 B7 V. Description of the invention (130) Ο

Among them, -AA-, -B_B_, R3, and R1 ^ R2〇 are as defined above, R25 is (^ to ^ alkyl.), And the 3_ position is subsequently deprotected by traditional hydrolysis and introduced again -Keto group and 5,7-hemiacetal, yielding a further intermediate hydrazone according to formula 209

[209] Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economics where-A-A ·, -B-B_ * R3 are as defined above. The epoxy portion of 9,11 is then introduced according to any method described above for converting a compound of formula II to a compound of formula I. Under the oxidizing conditions of the epoxidation reaction, the hemicondensation part is converted into 5,7-dilactone, thereby generating a further intermediate that meets the formula 2 n: CNS) A4 specification (210X297 mm) 593305 A7 B7 V. Description of the invention (ο

[211] As defined above. Any residues in which Α-Α-, -Β-Β-, and r3 are as follows:

(Please read the precautions on the back before filling in this page. J • Order · Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs [210] ^ 中 · Α_Α ·, _B_B ^ R3 As stated above, the traditional Wanfa oxidation method is 2ir Finally, the method described in Scheme 8 was used to convert 5,7. 2 to 7α-oxooxycarbonyl compounds, the intermediate of formula 2U was converted to epoxy mexrenol or other magic compounds. Therefore, the process of the complete plan 10 is as follows, and it is understood that at least the following steps can be particularly redundant without the need to recover intermediates. The complete method for the synthesis of plan i 0 is as follows: -134--^ 1 No, take Manpower applies Chinese National Standard (CNS) A4 specification (210X 297 mm) 593305 A7 B7 V. Description of invention (l32)

OR 'V /// CHO 丨 203】

Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

211J 0 Epile Leno -135- (Please read the notes on the back before filling this page)

This paper size applies to China National Standard (CNS) A4 (2I0X297 mm)

XXXIX Printed by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 593305 Invention Description (l33 As in the case of Plan 9 ψ ,,, 〖'the above-mentioned response to Plan 10 provides important, especially economic, processes; But Plan 10 is novel: it also has more common enzymes that are different from those specifically described. For example, 7-methylamido is introduced into 3-enol ether steroids to protect the 7-formamidine. _ △ s .j 〇, 6_3,4-enol ether, hydrolyzed 5,7-hemiacetal, and deprotection effect of subsequent performance 仏 * more effective, can be defined at the 17-position as R8 and R-substituted steroids, or Cheng Du + Kou 4 more specifically at the 17-position, are substituted by X′γ1, γ and C (17) as defined above: 7) ΠΙΜ (CHfc) 2. C Operating on steroids. Ten paintings] Another method of 10, including the use of various intermediates in the above range from the formula A 2 0 3 J A 2 1 0, respectively. Each intermediate from formula 03 to A2i i is a novel compound and can be used to prepare% oxmex Reynolds or other compounds of formula * ia according to the synthetic method of plan 10. In a specific embodiment, the complete method of planning the 10 process is as follows: -136 · This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm (please read the precautions on the back before filling this page)

593305 A7 B7 V. Description of the Invention (I34)

Printed by the Staff Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs -137 · (Please read the precautions on the back before filling this page)

This paper size applies the Chinese National Standard (CNS) A4 specification (210X 297 mm). Among the several plans explained above, γ — a reactionary method used in the method of the present invention, in the manufacture of epoxy US $ j clothing Ruox Lei Renuo and related compounds, to provide two-time softening. Key features include, in particular: the biotransformation of enzymes such as Conreno, Tada's monoketone or sterol, which are converted to &quot; ... Ge 9 derivatives Temple converts cold_sitosterol to 1 7 -keto-condensation); (1 &gt;) by containing 11 (^-or 9 ^ /, &gt;-|, / 丨 human 11 ay α-fluorenyl &lt; compound Dehydration leads to Λ 9,1 1 double bond ', and then through the oxidation of the static bond ΑΑ &quot;,,! 1 double bond to introduce epoxy groups; (C) By forming a shaft, the amine is hydrolyzed to H The diketone is reacted with the aging metal alkoxide to attach ^ oxoyl; ((1) 20-spiro dislocation is formed at the 17 position; (e) 5,7_dilactone is formed, and the Lactones are converted to 7-oxooxycarbonyl groups; ⑴ protects 3-ketones (including the formation of 2.0 at the "position" by converting to 3-alcohol ether or 3 'ketal during various conversion periods at other positions. Spiro oxygen burn ring). With a few restrictions, the four constituent process elements (b) to ⑷ 'can be operated in almost any order. Process elements ⑷ and (f) provide comparable flexibility. They provide formation Epoxy Mex Renault and The route of his formula 1 compound is simpler than the method of U.S. Patent No. 4,559,332-Midnight. In addition, they provide significant advantages in productivity and yield. In the reaction plan descriptions described above, the recovery of reaction products, Separating the consumer cooperatives from the Central Bureau of Standards of the Ministry of Economic Affairs and the consumer cooperatives can usually be accomplished by methods familiar to the artist. Unless otherwise stated, "The conditions, solvents, and reagents are traditional and there are no strict restrictions." Or both. However, 'Some special procedures, as expected above', provide favorable total yields and / or productivity, and / or intermediates and finals, which contribute to the various method steps and method planning. The advantages of the high quality of epoxy steroid products. -138- This paper size is suitable for the standard _ house standard (cm) "grid (2H) X297 mm) printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 593305 A7 ------- _B7 V. The practicality of the 20-spiroxane compound produced according to the present invention is described in

Grob U.S. Patent No. 4,559,332 is specifically incorporated herein by reference. The 20-spiroxane compound produced according to the present invention is well-known for its good biological properties, and is therefore a valuable hafnium-containing pharmaceutically active ingredient. For example, they have a strong aldosterone antagonist effect, so they reduce and use excessively high sodium retention and potassium excretion normalization due to aldosterone. Therefore, they have important therapeutic uses as diuretics for retention, such as in the treatment of hyperglycemia, cardiac insufficiency, or liver cirrhosis. 2-spiroxane derivatives are known to have the effect of aldosterone antagonists, see for example Fieser and Fieser: Steroids; page 708 (Reinhold Publ Corp., New York, 1959) and British Patent Application No. 1041,534 ; 17 /?-Hydroxy-2 1 -carboxylic acids and salts thereof having a similar effect are also known; see, for example, US Patent No. 3,849,404. Such compounds have so far been used therapeutically, however, they have a considerable disadvantage in that they always have certain and specific activities, and have troublesome results during or after routine long-term treatment. Particularly undesirable is the troublesome effect caused by the anti-androgen activity of known anti-aldosterone products. The following examples further illustrate the methods, processes, and compositions of the present invention, as well as the conditions and reagents used herein. Example 1 Using the growth medium listed in Table 1 to prepare the slanted surface-139- This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) (Please read the precautions on the back before filling this page) Order-^ 1. ^ 3305 A7 B7 Five λ invention description (l37) Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs Table 1 _YPDA (medium for slanted and flat surfaces) yeast extract raw sugar distilled water amount to -pH 6.7: use Ρ3ρ〇41〇% weight / volume adjustment to pH 5 minutes---About the bevel: 7.5 ml in a 18 χ 18 mm test tube-About the plane (10 cm 0): 25 ml in a 200 x 20 mm test tube Sterilize at 120 ° C for 20 minutes. PH 之后 after sterilization: 5 To produce the first-generation culture, distill the water (2 ml) in the seedlings of Scutellaria luteus; and 5 ml aliquots of the suspension = The tube is on each bevel as prepared above. Place the bevel at 25. The appearance of the surface of the culture after incubation at 0 ° was a white cotton-like mycelium. The back is tinted, the lower part is orange, and the upper part is yellow-orange. The first-generation slant culture was suspended in a sterile solution (4 ml) containing a Tween 80 nonionic surfactant (3% by weight), and an 0.15 ml aliquot of the suspension was used Come to inoculate 20 grams of Moo liters prepared with the growth media listed in Table 2 (please read the precautions on the back before filling out this page)-Order ^ 1. -140- 593305 A7 B7, Description of the Invention (138)- — Second generation bevel. Table 2 (for the second generation and the example bevel) Malt extract 20 g gland 1 g ~ 'glucose 20 g agar 20 g distilled water amount to -pH 5 · 3 Dispense into test tube (180x8 mm) 7.5 Ml-1000 ml sterilized at 120 ° C for 20 minutes (please read the precautions on the back before filling out this page) noodles · order printed by the Ministry of Economic Affairs Central Standard Bureau employee consumer cooperative is brown orange. Preparations are shown in Table 3. Protected medium with the composition listed in Table 3-Protected medium skim milk distilled water in a 250 ml flask containing 100 ml of distilled water was added to skim milk. Sterilize at 120 ° C for 15 minutes. Cool to 33 C 'and Use more than a day ago._ Suspend the cultures from the five generations of bevels in a 100 亳 ^ protective solution (15 亳 L). Divide the suspension in equal portions (each 0.5 各 L) Eight mixes are cold-dried in test tubes of ΙΟΟχΙΟ φ meters. These 141-paper sizes apply to Chinese National Standard (CNS) A4 (210X29? Mm) at _70 ° C to 593305. Consumption by employees of the Central Bureau of Standards Cooperative printed A7 --- _--- B7__V. Description of the invention (139) '~-Work cold in acetone / dry ice bath; East 2 () minutes, and then immediately move to pre-cooling; East to -4 (TC to -5 (TC drying chamber. Will Pre-cooled aliquots were at the micro: remnant pressure of the column and cooled at S-3 (rc; dry at the east. At the end of the cold-drying, add two to three sterile silicone particles with a humidity indicator to each Each test tube is sealed with a flame. To obtain a slant of the mother culture suitable for large-scale fermentation, aliquots of the freeze-dried culture prepared by the above method are suspended in distilled water (U ml), and 0.15 ml is used as In one aliquot, the suspension was used to inoculate a slant with a growth medium having the composition listed in Table 2. The mother slant was cultured at 25 ° C for 7 days. At the end of the culture, the culture developed on the slant was stored in 4. (: Bottom.) To prepare a conventional slant culture, the culture obtained from the mother slant is suspended in a sterile solution (4 ml) containing Tween 80 (3% by weight), and 0.15 ml is used as In one aliquot, distribute the resulting suspension to a coating that has been applied using the growth medium described in Table 2 Can be used to inoculate the initial seed flask for laboratory or industrial fermentation. To prepare the initial seed flask culture, remove the culture obtained from the conventional bevel preparation as described above and suspend In a solution (10 liters) containing Tween 80 (3% by weight), an aliquot of the resulting suspension, 0.01 ml, was introduced into a 500 ml fold containing a growth medium having the composition listed in Table 4. Flow flask 0 -142- The paper size is suitable for Guancai County (CNS) A4 size (21GX297 male diamond) — (Please read the precautions on the back before filling this page)

V. Description of the invention (14G Table 4 (for initial and transformation flask cultures and round-bottomed flasks) glucose 20 g gland 20 g yeast autolysate 20 g distilled water appropriate amount-pH 5.2-adjusted to 20% NaOH pH 5.8-Dispensed into a 500 ml baffled flask 100 ml-Dispensed into a 2000 ml round bottom flask (3 baffles) 500 ml-Sterilized 120 ° C x 20 minutes-pH sterilized after sterilization ~ 5.7 1 000 ml (Read the precautions on the back before you fill in this page.) Printed by the Consumer Cooperatives of the Central Standards Bureau, Ministry of Economic Affairs, at 28 ° C, by cultivating seed flasks on a rotary stirrer (200 rpm, 5 cm displacement) for 24 hours. A culture in the form of a mycelium-like mycelium having a diameter of 3 to 4 mm was produced. When observed under a microscope, the seed culture was found to be a pure culture that grew symmetrically with large fully entangled mycelium. Suspension The pH of the solution was 5.4 to 5.6. The PMV was determined to be 5 to 8% (3 000 rpm x 5 minutes) by centrifugation. By inoculating the second pellet (1 ml) from the seed culture flask on the second The 5 0 0 liter stirrer flask has the items listed in Table 4. The composition of the growth medium (100 liters) was used to prepare a transformation flask culture. The resulting mixture was cultured at 28 ° C for 18 hours in a rotary stirrer (200 rpm, 5 cm displacement). Check the culture It was found to include small spherical mycelium with a diameter of 3-4 mm. Upon microscopic examination, the culture was determined to be a pure culture with symmetrical and filamentous growth, in which the apical cells were filled with cytoplasm, while the older cells were almost There is no cavitation. The p Η 値 of the culture suspension is 5 to 5 · 2, which is determined by centrifugation -143 · This paper size applies the Chinese National Standard (CNS) A4 specification (210X 297 mm) 593305 A7 B7 V. Invention Note (PM) that PMV is between 10% and 15%. Therefore, the culture is considered suitable for converting Conranow to 11α-hydroxy Conranow. Reducing Conranow (1 g) to a particle size of approximately 5 microns, And suspended in sterile water (20 ml). To this suspension were added: 40% (w / v) sterile glucose solution; 16% (w / v) sterile autolyzed yeast solution; and sterile Antibiotic solution; The ratios indicated by the 0-hour reaction time in Table 5. By dissolving conomycin sulfate (40 mg), tetracycline HC1 (40 g), and cephalosporin (200 mg) in water (100 mg). Ml) to prepare antibiotic solution. Gradually add steroid suspension, glucose solution and autolyzed p master solution to the culture contained in the stirring flask. Table 5 During the biotransformation of Conreno in the stirring flask, specify Adding solids> Valley liquid (additives and antibiotics) C Please read the notes on the back before filling in this page j 丨 «· Printed by the Staff Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

During the reverse process, the reaction mixture was regularly analyzed to determine the glucose content. This paper rule Money 144-593305 A7 B7 V. Description of the invention (142) and determined by thin layer chromatography 11 Conversion. Additional Conranox enzymes and nutrients were added to the fermentation reaction mixture at a controlled rate during the reaction to maintain the glucose content in the range of about 0.1% by weight. The schedules for the addition of steroid suspensions, glucose solutions, autolyzed yeast solutions and antibiotic drops are listed in Table 5. The conversion reaction was continued on a rotary stirrer (200 rpm, 5 cm displacement) at 25 for 96 hours. During the fermentation period, 卩 11 卩 Fanyuan was between 4.5 and 6. as long as? ] ^ ¥ was raised to 600 / () or more, and 10 liters of broth culture was retrieved and replaced with 1000 ml of distilled water. During the reaction period, monitor the Conreno by sampling the broth at intervals of 4, 7, 23, 31, 47, 55, 71, 80, and 96 hours after the start of the reaction cycle, and analyze the sample by Ding ^. Disappeared and the appearance of n-Hydro Conreno. Determined from the progress of the reaction of these samples listed in Table 6. Table 6 ^-Bioconversion effect of Yicono Renuo in a stirred flask ^ time advance, time ~ conversion rate 7 f. = 0.81 基 f basis (please read the precautions on the back before filling this page). Order economy Printed by the Consumer Standards Cooperative of the Ministry of Standards

-145 · This paper size is in accordance with Chinese National Standard (CNS) A4 (21 OX 297 mm) 593305 A7 B7 V. Description of the Invention (I43) Example 2 The initial seed flask culture was prepared by a method similar to that described in Example 1. Thing. A nutritional mixture was prepared having the composition listed in Table 7. Table 7 Amount of transformation culture used in a 10 liter glass fermenter g / L glucose 80 g 20 gland 80 g 20 yeast autolysate 80 g 20 antifoam agent SAG 47 1 0.5 g deionized water amount to-at 130 Sterilize the empty fermenter for 30 minutes at ° C • Fill with 3 liters of deionized water and heat to 40 ° C-Add and stir the ingredients of the culture medium-Stir for 15 minutes with a volume of 3.9 liters-pH 値 is 5.1 -Use NaOH 20% weight / volume to adjust to 5.8-Sterilize at 120 ° C x 20 minutes-pH 后 after sterilization is 5.5-5.7 4 liters printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back first) (Fill in this page) The initial loading of the nutrient mixture (4 liters) into a 10 liter geometric volume conversion fermenter. The fermenter is a cylindrical centrifuge tube with a length to diameter ratio of 2.58. Provide 400 r p m turbo agitator: agitator with two No. 2 wheel discs, each with 6 blades. The outer diameter of the impeller is 80 mm, the radial diameter of each blade is 25 mm, and the height is 30 mm. The upper wheel is located below the top of the container. -146- This paper size applies to Chinese National Standard (CNS) A4 (210X 297) (Mm) 593305 A7 B7 V. Description of the invention (I44) 280 mm, the lower wheel is 36.5 mm below the top, the container is folded 210 mm, and the inner wall of the container extends 25 mm radially inward. Ma also mixed the seed culture (40 ml) with the nutrients contained in the fermenter, and cultivated it for 22 hours at a pressure rate of 0.5 kg / cm2 under a pressure of 0.5 kg / cm2, Establish transformation cultures. in. At hours, the culture had a PMV of 20-25% and a pH of 5 to 52. A suspension containing Conreno (80 g) in sterile water (400 ml) was prepared and a 10 liter portion was added to the mixture in a conversion fermenter. At the same time, in the proportion indicated in Table 8 at a reaction time of 0 hours, add 40% (weight I / vol) sterile glucose solution, 16% (weight / vol) sterile autolyzed yeast solution and sterile antibiotic solution. . An antibiotic solution was prepared in a similar manner as described in Example 1. Table 8 During the bioconversion process of Conreno in a 10 liter glass fermenter, the specified addition of steroids and solutions (additives and antibiotics) reaction time is small. The steroid suspension is lifted to about a gram of glucose solution, ml of autolyzed yeast solution, and antibiotic solution. 0 10 4 25 12.5 40 4 25 12.5 8 10 4 25 12.5 12 25 12.5 16 10 4 25 12.5 20 25 12.5 24 10 4 25 12.5 40 -147- The paper size is suitable for financial use_CNS A4 size (2iGx297 Public Fantasy (Please read the notes on the back before filling this page)

Employees' Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5 3 3 9 5 A7 B7 Legislation and Description of Inventions (145) 28 32 36 40 44 48 52 56 60 64 68 72 76 10 4 25 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 25 25 12.5 12.5 4 25 25 25 25 25 25 25 25 25 25 25 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 (please read the notes on the back before filling this page) Printed by Cooperative Society 80 84 88 As the reaction proceeds, the reaction mixture is periodically analyzed to determine the glucose content, and the conversion of 11-hydroxy-Conorno is determined by thin-layer chromatography. Based on the TLC analysis of the reaction broth samples described below, additional Conorno was added to the reaction mixture in accordance with the consumed Conorex enzyme. The glucose content was also monitored, and when the glucose concentration dropped to about 0.055% by weight or less, a supplementary glucose solution was added to raise the concentration to about 0.25% by weight / 0. Nutrition and antibiotics are also added at different times in the reaction cycle. Addition plan of steroid suspension, glucose solution, autolyzed yeast solution and antibiotic solution -148- Two paper scales are applicable to China National Standards (CNS) A4 specifications (21 × M? Publication) Central Standards Bureau of the Ministry of Economic Affairs-industrial consumption Printed by the cooperative 593305 A7 ____ B7 5. The description of the invention (I46) is shown in Table 8. The conversion reaction was continued for 90 hours under a positive top pressure of 0.3 kg / cm² at 0.5 parts by volume of air (vvm) per part of liquid per minute. The temperature was maintained at 28 ° C until pv Μ reached 45%, then decreased to 2 ° C and maintained at this temperature, so that ρ ν μ grew from 45 to 60%, and then controlled at 24 ° C under. The initial agitation rate was 400 rpm and increased to 700 r p m after 40 hours. The pH was maintained between 4.7 and 5.3 by adding 2 μ orthoacetic acid or 2 M NaOH as indicated. When foaming develops, foaming is controlled by adding a few drops of antifoam agent SAG 47 1. During the reaction, the loss of Conreno and the appearance of 1 1 α-hydroxyconreno were monitored by TLC analysis of broth samples at 4-hour intervals. When most of the Conorno disappear from the broth, add additional amounts. The reaction was stopped after all the Conranow was added. At this time, TL c analysis showed that the concentration of Conranox enzymatic substance relative to 1 1 -hydroxy Conranow had been reduced to approximately 5%. Broth, the mycelia are separated from both liquid meat. The mycelium part was resuspended in ethyl acetate, using approximately 65 parts by volume (5.2 liters) per gram of Conreno loaded during the reaction. The mycelium suspension in ethyl acetate was refluxed for 1 hour under agitation, cooled to about 20 C 'and filtered on a porcelain funnel (Buchner). The mycelium cake was continuously rinsed with ethyl acetate (five volumes; 0.4 liters in mother Concan); and deionized water (500 ml) to remove the ethyl acetate extract from the filter cake. Discard the filter cake. The extract-rich solvent wash and water wash were collected in a separator, and then allowed to stand for 2 hours to separate the phases. Then discard the liquid phase and concentrate the organic phase under empty space to the remaining volume of 35-149- This paper size applies Chinese National Standard (CNS) A4 specification (210X297 mm) (Please read the precautions on the back before filling this page ) Order- ^ 1. Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs 593305 A7 B7 V. Description of Invention (I47) ml. Allow the bottom to cool to 15 ° C and keep stirring for about 1 hour. The suspension of the residue was filtered to remove the crystallized product, and the filter cake was rinsed with ethyl acetate (40 ml). After dehydration, it was determined that the amount of 1 1 α-Turkey Conreno produced was 60 g. Example 3 A spore suspension was prepared from a routine bevel in the manner described in Example 1. In a 2000 ml baffled round bottom flask (3 baffles, 50 mm x 30 mm each), an aliquot (0.5 ml) of the spore suspension was introduced into a nutrient solution having the composition listed in Table 4 ( 500 ml). The resulting mixture was cultured in a flask for 24 hours at 25 ° F. on another stirrer (1 200 stroke per minute; displacement 5), thereby producing a culture, which was clearly observed during microscopy. It is a pure culture with fully tangled mycelium. The ρ Η 値 of the culture is between approximately 5.3 and 5.5, and the PMV (determined by centrifugation at 3,000 rpm for 5 minutes) is 8 to 10%. Using the culture thus prepared, a seed culture was prepared in a stainless steel fermenter having a vertical cylindrical centrifuge tube having a geometric volume of 600 liters and an aspect ratio of 2.31 (length = 9 85 mm, diameter = 425 mm). A disc-turbine stirrer having two discs was provided, each disc having six blades each having an outer diameter of 240 mm and each blade having a radial diameter of 80 mm and a height of 50 mm. The upper wheel is at a depth of 780 mm from the top of the fermenter, and the second wheel is at a depth of 995 mm. The vertical baffle has a length of 89 mm and extends radially inward from the vertical wall in the fermenter by 40 mm. The agitator was operated at 170 rpm. Introduce the nutrient mixture (丨 00 liters) with the composition listed in Table 9 into the fermenter 'and then introduce a part of the pre-150 prepared above with a pH of 5.7- Standard (CNS) six 4 specifications (21〇χ 297 mm)-(Please read the precautions on the back before filling out this page)-Order 593305 A7 B7 V. Description of the invention (148) Inoculum (1 liter). Table 9 For the cultivation of plants in a 160 liter fermenter, about 8 liters of seeds are needed to produce a fermenter. G / liter glucose 2 kg 20 glands 2 kg 20 yeast autolysate 2 kg 20 antifoam agent SAG 471 0.010 kg trace Deionized water is appropriate to-sterilize the empty fermenter at 130 ° C for 30 minutes-fill 6 liters of deionized water and heat to 40 ° C-add and stir the ingredients of the culture medium-stir for 15 minutes with 95 liters Volume-Sterilization at 121 ° C for 30 minutes-pH 后 after sterilization-5.7-Add sterile deionized water to 100 liters 100 liters. The cooperative prints to incubate the inoculated mixture at a 0.5 l / l · min aeration rate under a 0.5 kg / cm2 surface pressure for 22 hours. The temperature was controlled at 28 ° C until PM V reached 25%, and then decreased to 25 ° C. pH 値 is controlled in the range of 5 -1 to 5.3. The growth of the lean body volume is shown in Table 10 along with the pΗ 値 and dissolved oxygen content of the seed culture reaction. -151- This paper size applies to Chinese National Standard (CNS) Α4 size (210 × 297 mm) 593305

7 7 AB V. Description of the invention (i49) Table 1 0 Time course of mycelium growth during seed culture fermentation. Hour pH during fermentation. Filled mycelium volume (PMV)% (3000 rpm, 5 minutes). 0 5 · 7 ± 0 · 1 100 4 5 · 7 soil 0.1 100 8 5.7 soil 0 · 1 12 soil 3 85 soil 5 12 5.7 soil 0.1 15 soil 3 72 soil 5 16 5.5 ± 0.1 25 soil 5 40 soil 5 20 5.4 Soil 0.1 30 Soil 5 35 Soil 5 22 5.3 Soil 0.1 33 Soil 5 30 Soil 5 24 5.2 Soil 0.1 35 Soil 5 25 Soil 5 (Please read the notes on the back before filling out this page) Printed by the Employees' Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs The seed culture thus prepared was used to complete the transformation fermentation process in a vertical cylindrical stainless steel fermenter with a geometric volume of 1.0 2 meters in diameter, 1.5 meters in height, and 1.4 cubic meters in geometry. A fermenter was provided with a turbo agitator with two propellers, one 867 cm below the top of the reactor and one 4 3 5 cm below the top. Each wheel has six blades each with a radial diameter of 95 cm and a height of 75 cm. The vertical baffle is 1440 cm high and extends 100 cm radially inward from the vertical wall in the reactor. A nutritional mixture having the composition listed in Table 11 was prepared. -152- This paper size applies to China National Standard (CNS) A4 (210X297 mm) 593305

A

7 B V. Description of the invention (15) Table 1 1 Biotransformation culture used in 1000 liter fermenter g / L glucose 16 kg 23 gland 16 kg 23 yeast autolysate 16 kg 23 antifoam agent SAG 471 0.080 kg of trace deionized water is appropriate to-sterilize the empty fermenter at 130 ° C for 1 hour-fill 600 liters of deionized water and heat to 40 ° C-add and stir off the ingredients of the culture medium-stir for 15 minutes, It has a volume of 650 liters-sterilized at 121 ° C for 30 minutes-the pH after sterilization is 5.7-add sterile deionized water to 700 liters and 700 liters! | J --------— (please first 1 Read the notes on the back and fill in this page again. 1T introduce the nutritional mixture (pH = 5.7) with the initial loading (700 liters) into the fermenter, and then introduce the seed inoculum (7 liters) of this experiment prepared as described above. The Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs prints the nutrient mixture containing the inoculum at an inflation rate of 0.5 liters / liter-minute at a surface pressure of 0.5 kg / cm2 for 24 hours. The temperature is controlled at 28 ° C, and the stirring rate is 1 10 rpm. Shown in Table 12 The growth of mycelial volume, together with the pΗ 値 and dissolved oxygen content of the seed culture reaction. -153- This paper size applies the Chinese National Standard (CNS) A4 specification (210 × 297 mm) 593305 V. Description of the invention (151) Table 12 Time course of mycelium growth in a fermenter for transformation culture. Hours pH during fermentation. Filling of mycelium volume (PMV)% (3000 rpm X 5 minutes). Dissolved oxygen% 0 5.6 soil 0.2 100 4 5.5 soil 0 · 2 100 8 5 · 5 soil 0.2 12 soil 3 95 ± 5 12 15 soil 3 90 soil 5 16 5.4 soil 0.1 20 soil 5 75 ± 5 20 5.3 soil 0.1 25 soil 5 60 soil 5 22 5.2 soil 0.1 30 soil 5 40 soil 5 ( Read the notes on the back before filling in this page.) At the end of the culture, small pellets of mycelium are obtained, but the pellets are usually small and fairly loose. The scattered mycelium is suspended in the broth. Finally The pH 値 is 5.1 to 5.3. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs, the transformation culture thus produced is added to Conreno (1.2 5 kg in sterile water (5 liters); micronized into 5 Μm) suspension. The proportions specified in Table 14 for reaction time 0 Add the additive solution and antibiotic solution. The composition of the additive solution is listed in Table 1 3. -154- This paper size is applicable to Chinese National Standard (CNS) A4 specification (210X297 mm) 593305 A7 B7 V. Description of the invention (i52 ) Table 1 3 Additive solution (for biotransformation cultures) Amount of dextrose 40 kg yeast autolysate 8 kg antifoam agent SAG 4 7 1 0 · 0 1 0 kg Deionized water amount to-at 130 ° Sterilize 150 liters of empty fermenter under C 丨 hour-fill 70 liters of deionized water and heat to 40 ° C-add and mix, the ingredients of the "additive solution"-good: drop for 30 minutes, with 95 liters Volume-pH 値 is 4.9-Sterilize at 120 ° C for 20 minutes-pH 后 after sterilization is 5 100 liters (please read the precautions on the back before filling out this page) Order printed by the Staff Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs Using a 0.5 l / litre-minute inflation rate and a surface pressure of 0.5 kg / cm², the biotransformation is completed in about 96 hours, and the range of ρΗ 値 is between 4.7 and 5 · 3 'according to It needs to be adjusted by adding 7 $ μ N a 〇 Η or 4Μ ^^ 〇4The stirring rate was initially 100 rpm, increased to 165 rpm at 40 hours, and 250 rpm at 64 hours. The initial temperature was 28 ° F, dropped to 26 ° C when PMV reached 45%, and dropped to 24τ when PMV rose to 60%. When you need to control foaming, use Drop 471. The glucose content during the fermentation was monitored at 4-hour intervals. When the glucose concentration was below 1 gpl, a supplemental amount of sterile additive solution (10 liters) was added. The disappearance of Conorno and -155- was also monitored by HPLC during the reaction.

, I--I--593305 A7 ------------- B7 _ V. Description of the invention (153) 1 1 Pan-via the emergence of Gicono Reynolds. When the initial Conorno load of at least 9 0/0 has been changed to 11 Shen Fafa Conreno, add 2.5 kg of Conreno supplement. When 90% of the supplemented Reynolds showed a shift, another 1.250 kg of replenishment was introduced. Add more replenishment (1.25 kg each) using the same standards until the total reactor load (20 kg) has been introduced. After all of the Reynolds in Shao have been charged into the reactor, the reaction is stopped. At this time, the concentration of the unreacted Conreno is 5% with respect to the amount of 1-α-hydroxyconreno produced. The schedule for the addition of Conranox, sterile additive solutions, and antibiotic solutions is shown in Table 14. Conranol J and Antibiotics: Table 14 During the conversion of the branch, add steroids and solutions to the fermenter (add * :). The reaction time is small. Liter volume approx. 0 1.250 1.25 10 8 700 4 10 8 1.250 2.5 10 12 10 16 1.250 10 20 10 24 1.250 5 10 8 800 28 1.250 10 32 1.250 10 36 1.250 10 40 1.250 10 44 1.250 10 48 1.250 12.5 10 8 900 Printed by the Consumers' Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the notes on the back before filling this page) -156 · This paper size applies to China National Standards (CNS) Α4 specifications (210 × 297 mm) Printed by the cooperative 593305 Α7 ________ Β7 V. Description of the invention (I54) 52 1.250 10 56 1.250 10 60 1.250 10 64 1.250 10 68 1.250 10 72 1.250 20 10 8 1050 76 0 80 84 88 92 Total When the biotransformation is completed, Centrifuge in a basket centrifuge tube to isolate mycelia from the broth. By HPLC, it was determined that the filtrate contained only 2% of the total α-hydroxyconorno in the harvest broth, and was therefore excluded. The mycelium green was floated in ethyl acetate (1000 liters) in an extraction tank with a capacity of 2 m3. The suspension was heated under reflux and ethyl acetate under reflux for one hour, then cooled and centrifuged in a basket centrifuge tube. Rinse the mycelium cake with ethyl acetate (200 liters) and discard it. The steroid-enriched lotion extract was allowed to stand for one hour to separate the aqueous phase. The aqueous phase was extracted with a larger amount of ethyl acetate (200 liters), and then the solvent phase was clarified by throwing π ~ CO 贫 ~~, and placed in a concentrator (500 liter geometric volume), j = It was concentrated to a residual volume of 00 liters. When performing evaporation, the mixed extract and rinse solution of the concentrator was 100 liters at first. When the solvent was removed, the volume was maintained constant by adding the mixed solution continuously or periodically. After completing the action steps, cool to the bottom, which is still at the bottom, and then (please read the precautions on the back before filling this page), 1T -157-

593305 7 V. Description of the invention (155)-^, then filter on a porcelain funnel filter. Rinse the pot of the concentrator with ethyl acetate (20 liters), and then use the rinse solution to rinse the cake on the filter. The product was dehydrated at 50 ° C. for 16 hours under vacuum. 14 kg of η-Hydroxy-Connor is produced. Example 4 Freeze-dried spores of AMANRRL 405 were suspended in a corn slurry growth medium (2 wells) having the composition listed in Table 15: ---------- one hour / work, VL seven Τ: Table 15-Clay slurry culture medium (for the growth of the original seed culture, its corn slurry 30 grams of yeast extract, 15 grams of monovalent brick acid, 3 grams of glucose (filled after sterilization), distilled water amount to 1000 亳The pH was increased to 4.6, adjusted with 20% NaOH, 50 ml was dispensed into a 250 ml conical flask, and sterilized at 121 ° C for 20 minutes and 30 g (please read the precautions on the back before filling this page) 丨 « · Ordered by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs for the propagation of spores, the resulting suspension was used to inoculate on agar culture plates. Ten agar culture plates were prepared, each with the composition listed in Table 16. Solid glucose / yeast extract / phosphate / agar growth medium: Table 16-GYPA (glucose / yeast extract / gallate / agar culture plate) glucose (filled after sterilization) 10 g yeast extract 2.5 g k2hpo4 3 g agar 20 The amount of steamed noodles is 1000ml -158- This paper size is applicable to Chinese National Standard (CNS) A4 (210X297mm) * -------: ',》 ~ 一 #gangsha service : Table 17-GYP / Glycerin (glucose / yeast extract for raw material vials / gallate / glycerol ecstasy glucose (filled after sterilization) 10 g yeast extract 2.5 g K2HP〇3 3 gϊΐ ~~ '&quot;-Appropriate amount of distilled water to 100 ml 20 grams of paper ____ Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 593305 A7 ----------- B7 V. Description of the invention (I56 ) Adjust p Η 値 to 6.5 Sterilize for 30 minutes at 121 ° C. Move 0.2 ml aliquots of suspension to the surface of each culture plate. Incubate the culture plate at 2 $ C for ten days Then, spores harvested from all culture plates were sterilized at 121 C for 30 minutes. __ The suspension was divided into twelve bottles, each vial was transferred into 丨 ml. These wash bottles constitute the main cell bank, which can be removed Generate a working cell bank to generate Conranow's inoculum for conversion to n-Hydroxy Conranow. Primarily = Cell Bank The vial was stored in the vapor phase of a liquid nitrogen refrigerator at 13 ° C. You began to prepare a working cell bank, re-suspending the spores obtained from a major cell bank vial = aseptic growth with the composition listed in Table 15. The suspension was divided into ten 0.2 ml aliquots, and each aliquot was used to inoculate an agar culture plate having the composition listed in Table 16 with a solid growth medium. These plates were cultured at 25 ×: T for 10 days. After three days of incubation, the unwanted growth medium was brown-orange. A large number of golden yellow spores are produced during cultivation. Spores were harvested from each dish using the procedures described above for preparing major cell banks. Prepared in total (degree applicable towel S _ 辟 (CNS)

-159 · 593305 A7 ________B7_ 5. Description of the Invention (I57) 100 vials, each containing 1 ml of suspension. These vials constitute a working cell bank. Also by storing in _ 丨 3 0. (: The working cell bank vial was stored in the vapor phase of a liquid nitrogen refrigerator. The growth medium (50 ml) with the composition listed in Table 15 was placed in a 250 ml conical flask. (0.5 ml) working cell suspension was introduced into the flask and mixed with growth medium. The inoculation mixture was incubated at 25 ° C for 24 hours, yielding an original with a mycelium volume percentage of approximately 45% Seed culture. When the culture was visually inspected, it was found to contain! _ 2 mm-diameter-like spherical mycelium; and when observed under a microscope, it was apparently a pure culture. The second-generation seed culture was cultivated by The growth medium having the composition listed in Table 5 was introduced into a 2-8 liter Fernbach flask and inoculated with a portion (10 ml) of the original seed culture of this experiment, which was prepared as described above. At 25 Incubate the inoculated mixture for 24 hours on a rotary stirrer (200 rpm, 5 cm displacement) at ° C. At the end of the cultivation, the culture showed the same characteristics as the original seed culture described above, and was suitable for use. Conranos Bio is transformed into 1 1 ^ # in the transformation fermentation of Conrad. Operating in the Braun Biostat fermenter, its structure is as follows: 15 liters, with a round bottom printed by the Consumers Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. (Please read the precautions on the back before filling in this page) South degree: 5 3 cm diameter: 20 cm H / D: 2.65 propeller: 7.46 cm diameter, 6 blades, each 2.2 magic. 4 cm advance interval : From the bucket; | Department 6 5 · 5, 1 4.5 and 2 5.5 cm -160- This paper size applies to the Chinese National Standard (CNS) A4 (210X297 mm ~ ---- 593305 A7 Table 1 8 (Fruit bucket 4 stand for biotransformation of rolls in a 10 liter fermenter) Amount / liter glucose 160 g 20 g gland 160 g 20 g yeast extract 160 g 20 g antifoam agent SAG 471 4.0 0.5ml Conrador deionized water to 7.5 liters Sterilized at 121 ° C for 30 minutes 160g 20g Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs ____ B7 V. Description of the invention (158) ^ Baffle plate: 4 1 · 9 X 4 8 cm sprayer · 1 · 1 cm diameter ' 21 holes ~ 1 mm diameter temperature control: Provided by means of an external sleeve to suspend Conreno in deionized water (4 liters) at a concentration of 20 g / l, and add one portion The growth medium of the listed composition, while stirring the mixture in the fermenter at 300 rpm. (Please read the precautions on the back before filling in the suspension on this page for 15 minutes, and then use additional deionized water to make Volume up to 7.5 liters. At this time, by adding 20 wt%. NaOH solution, the pH of the suspension was adjusted to 5.2 to 6.5, and then sterilized by heating in a Braim E fermenter to 1 2 1 ° C for 30 minutes. Ρ η 値 after sterilization was 6.3 soil 0.2, and the final volume was 7.0 liters. A portion (0.5 liter) of the second seed culture of this example prepared as described above was used to inoculate a sterile suspension, and the volume was up to 8.0 liters by adding a 50% sterile glucose solution. At 28. 〇 Fermentation is performed at a temperature of 50% until PMV reaches 50 ° C, and then drops to 26 ° C, -161- This paper size applies Chinese National Standard (CNS) A4 specifications (210X297 mm 1) Staff of the Central Bureau of Standards, Ministry of Economic Affairs Printed by the Consumer Cooperative 593305 A7 __________ ^ 5. Description of the Invention (159) and when the PMV exceeds 50%, it is reduced to 24% in order to maintain a fixed PMV of less than about 60%. Air is introduced through a sprayer based on the original liquid volume At a rate of 0.5 vvm and maintaining the pressure in the fermenter at 700 mm Hg. Start stirring at 600 rpm and gradually increase the dissolved oxygen content to 30% by volume as needed rpm. Monitor the glucose content. After the initial glucose concentration is reduced to 1% due to the consumption of the fermentation reaction, supplemental glucose is provided through a 50% by weight sterile glucose solution in order to maintain the residue in the batch cycle It has a concentration of 0.05% to between. The pH 前 before inoculation is 6.3 ± 0.2. After the pH 値 drops to about 5.3 at the beginning of the fermentation period, the residue of the cycle is added by adding ammonium hydroxide. The pH 値 was maintained in the range of 5 · 5 ± 0 · 2. Foaming was controlled by the addition of a solid polyethylene glycol antifoam provided by OSI Specialties, Inc. under the trade name s AG 4 7 i. Culture The growth of the material initially occurred during the first 24 hours of the cycle, at which time PMV was about 40%, the pH was about 5.6, and the amount of dissolved oxygen was about 50% by volume. Conreno started even when the culture was growing. During biotransformation, monitor the Conranow and n "_ Concord" by analyzing daily samples. The samples were extracted with hot ethyl acetate, and TLC and PLC were used. Analyze the resulting sample solution. When the residual Conranow concentration is approximately 10% of the original concentration, the bioconversion is considered complete. The approximate conversion time is 110 to 300 hours. When the bioconversion is complete At this time, the biological mass of the mycelium was separated from the broth by centrifugation. The supernatant was extracted with an equal volume of ethyl acetate, and the liquid layer was discarded. The portion of the mycelium was resuspended in ethyl acetate , Each gram is loaded with fermentation anti-162- This paper size applies to the Chinese standard (CNS) &amp; See Grid (297 mm factory (please read the notes on the back before filling this page) 1 «· Order 593305 Printed by the Employees' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs Α7 Β7 V. Description of the invention (16) The Reynolds of the reactor uses about 65 parts by volume. Under stirring, the mycelium suspension is refluxed for one hour, cooled to about 20X :, and filtered on a porcelain funnel. The perireone of the Reynolds is filled in the fermenter Five volumes of ethyl acetate were used to rinse the mycelium cake twice 'and then rinsed with deionized water (1 liter) to remove residual ethyl acetate. Mix aqueous extracts, rich solvents, solvent rinses, and water washes. Discard or re-extract the residual depleted mycelium filter cake, based on the analysis of the remaining steroids. Allow the mixed liquid phases to stand for two hours. The liquid phase was then separated and discarded, and the organic phase was concentrated under vacuum to a volume of about 500 liters. Then leave the bottom to cool to 15 ° C and slowly stir off for about an hour. The crystalline product was recovered by filtration, and rinsed with cold ethyl acetate (100 ml). The solvent was removed from the crystals by evaporation, and the crystallized product was dehydrated under a vacuum of 50 t. As described in Example 4, freeze-dried spores of A. flavus ATTI 1 85 00 were suspended in corn slurry growth medium (2 l). Ten agar plates were also prepared in the same manner as in Example 4. These plates were grown and harvested as described in Example 4 to provide the main cell bank. The vial containing the main cell bank was stored in the vapor phase of a liquid nitrogen refrigerator. The working cell bank was prepared from the vial of the main cell bank as described in Example 4 and stored at -130. (: Nitrogen refrigerator. The growth medium (300 ml) with the composition listed in Table 19 was charged into a 2 liter deflection flask. An aliquot (3 ml) of the working cell suspension was introduced into the flask. . On a rotary stirrer (200 rpm, 5 cm displacement), the inoculation mixture was grown at 28π for 20-24 hours to produce an original seed culture with a large size of -163-. This paper size applies to Chinese national standards (CNS ) Α4 specification (210X297mm) ~ &quot; -----. (Please read the notes on the back before filling out this page} 1 «. Order 593305 A7 ______B7 V. Description of the invention (161) Approximately 45% of the filling Mycelial volume percentage. When the culture was visually inspected, it was found to contain small spheroid-like mycelia with a diameter of 1 to 2 millimeters; and during microstatic examination, it was shown to be a pure culture. Table 19 Original And a second seed culture A certain amount per liter of glucose per liter (loaded after sterilization) 20 g gland 20 g yeast extract 20 g distilled water to 1000 ml sterilized at 12 1 ° C for 30 minutes (please read the back Please fill in this page again)) The second seed culture of the staff consumer cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs shall be used for the cultivation of the second seed culture. Start the incubator with 160 ml to 200 liters of the original seed culture of this example. Inoculate the fermenter. Prepare as described above. Incubate the inoculation mixture at 28 ° C for 18-20 hours at 200 rpm. The aeration rate was 0.5 vvm, and at the end of breeding, the culture showed the same characteristics as described above for the original seeds. The transformation was operated in a 60 liter fermenter, generally in the manner described in Example 4, except for growth The culture medium has the composition listed in Table 20, and the second seed culture taken from the initial load is 350 liters to 700 ml. The stirring rate is initially 2000 rp m, but because of the need to maintain the solubility Oxygen Above vol%, it increased to 500 rp m. The approximate bioconversion time for 20 grams / liter of Conorno is 80 to 160 hours. -164- The scale is applicable to China National Standard (CNS) A4 specifications ( 210X297 mm) &quot; '--- Order A7 B7 Five invention description (162 Table 20 Growth medium i Glucose (filled after sterilization) Gland

Yeast extract in the form of a 20% slurry in sterile water jZ ± jij ^ _35 liters, at 12rcT ^^ IT Example 6 Γ Please read the notes on the back first ¾ then fill out this page) 1 «Use from according to the examples A spore suspension of the working cell silver ten prepared by the method described in 4 was used to prepare the original and second seed cultures substantially also according to the method described in Example 4. Using the second Fengzi culture produced in this way, two Qin biotransformations were performed according to the type of modification method illustrated in Figure i, and two rounds were performed in the method illustrated in Figure 2. Table 2 Η lists the degree of transformation of the transforming growth medium, Conreno supplement schedule, and harvest time. The R2 Α process uses a Conreno addition plan based on the original shell similar to Example 3, while the R2 c process modifies instance 3 by adding only two; j Conreno, once at the beginning of the batch and another It's 4 after 24 hours. In the r2b and r2d process, the full name Conranow load is imported at the beginning of the batch and the pass is completed in the manner described in Example 4. Conranos is sterilized in a separate container and glucose is added during the process. Use a chapter 4 Waring blender to reduce lumps generated during sterilization. In R2 -165, this paper size applies Chinese National Standard (CNS) 8-4 specifications (210x 297 feet).

n n I 593305 A7 B7 5. In the process of the invention (163) and R2 B, the introduction of Conreno in a methanol solution into this batch of reactions is a process different from Examples 3 and 4 in this respect. Table 2 1-Description of the original Conranox biotransformation process Process number R2 A R2B R2C R2D medium (g / l) Corn slurry 30 and R2A process 30 and R2C process yeast extract 15 same 15 same NH4H2P04 4 3 glucose 15 30 OSA 0.5ml 0.5ml pH 値 2.5N NaOH 2.5N NaOH Adjusted to 6.0 Adjusted to 6.5 Conreno 10g / 80ml A 80g / 640 milliliter sterilized and blended; sterilized and mixed with alcohol, at 0, 1S, liters of methanol, added at 0 in 0 hours; 25 grams together at 0, 24, 30, 36, 42, hours; 200 50, 56, 62 and 68 added at 24 hours; Harvest time 143 hours 166 hours 125 hours 104 hours Biotransformation 45.9% 95.6% 98.1% 95.1% Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) In the process R2A and R2B, The methanol concentration accumulated to about 6.0% in the fermented beer and was found to inhibit the growth and biotransformation of the culture. However, based on the results of these processes, it is inferred that methanol or other water-miscible solvents can be effectively supplied at lower concentrations in order to increase Conorno's loading and provide it in the form of fine-grained precipitates, providing large The interface area is -166- This paper size applies to Chinese National Standard (CNS) A4 specification (210X297 mm) 593305 A7 __________ B7 V. Description of the invention (164) Printed by the Consumers' Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs makes Kang Lei Exposure to reaction. It was confirmed that Connor was stable at sterilization temperature (21 ° C), but converged into thick blocks. Using a Welling's blender, the agglomerates are extruded into fine granules to allow them to transform smoothly into products. Example 7 A spore suspension obtained from a working cell bank produced according to the method described in Example 4 was used, generally also in the manner described in Example 4, to produce: the original and the second seed culture. The description and results of Example 7 are shown in Table 22. Using the second seed culture produced in this way, a biotransformation (R3C) as described in Example 3 was performed in vivo, and three biotransformations according to the method generally described in Example 5 were performed. In the last three processes (R3A, R3B, and R3D), Conreno is sterilized in a hand bucket with growth media other than glucose. Supply unsterilized glucose from other buckets. The sterile Conreno suspension is introduced into the fermenter before inoculation or early in the biotransformation. In process R3] B, supplemented sowing conreno and growth medium were introduced at 465. The Conranow lumps formed during sterilization were removed via a Wehring blender, and the resulting fine-particle suspension was fed into the fermenter. The transformation growth medium Kangtian% addition plan, nutrient addition plan, harvest time, and degree of transformation of these processes are listed in Tables 22 and 23. 0 (Please read the precautions on the back before filling out this page j -Order &gt;-^ 1. -167- This paper size applies Chinese National Standard (CNS) A4 (210X297 mm) 593305 V. Description of Invention (i65) Printed by the Consumers' Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs Table 2 2-Conranos Biotransformation Process description Process number R3 A R3B R3C R3D medium (g / l) and process R3A corn slurry 30 and process R3A gland: 20 same yeast extract 15 same yeast extract: 2〇NH4H2P〇4 3 glucose: 20 glucose 15 OSA3 Ml of OSA 0.5 ml with 2.5 N NaOH pH 値 adjust with 2.5 N NaOH to 6.5 adjust to 6.5 Load Conranos Sterilize Conranos and blend it with Process R3A Non-sterilized Conrad and Process R3A. BL50 g with BI: 50 Knorr: By the same BL50 grams in Table 23 16.5 hours: 110 grams 16.5 hours: 110 grams Scheduled 16.5 hours: 110 46.5 hours: 80 grams per gram supply see 23 See Table 23 See Table 23 See Table 23 Harvest time 118.5 hours 118.5 hours 118.5 hours 73.5 hours Biotransformation 93.7% 94.7% 60.0% 68.0% (Please read the notes on the back before filling this page) -168- This paper size applies Chinese National Standard (CNS) A4 specification (210X297 mm) 593305 A7 B7 Five invention descriptions (I66)

Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 200g / 2 Glucose and Yeast Extracted Antibiotics 20mg Grammycin 20 mg tetracyclic long medium ginseng medium long gram g in 1 liter of water 100 mg cephalosporin see table 22 see table 22 see table 22 grammycin in 50 ml towels g / l g / l g / 0----50 g / 0.4 liter 50 g / 0.4 liter 50 g / 0.4 liter 14.5 16 100 25 50 ml--16.5 ~---110 g / 1.2 liter 110 g / 1.2 110 g A .2 liter Liter 20.5 16 140 25-—- 1 28.5 16 140 25-• ㈣ — 34.5 16 150 25 — __ — 40.5 16 150 25 50 ml —-46.5 880 130 25-80 g / 0.8 liter — 52.5 160 120 25 1 — -58.5 160 150 25-—-64.5 160 180 25 50 ml —-70.5 160 140 25 — —--(Please read the notes on the back before filling out this page) Order -169. This paper size applies to Chinese national standards ( CNS) A4 specification (21〇 &gt; &lt; 297 mm) 593305 A7 B7 Note (I67) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs Due to the horse-like growth, high-viscosity fermented broth was seen in all four processes in this example. In order to overcome the obstacles caused by high viscosity caused by aeration, mixing, pH control and temperature control, it is necessary to increase the aeration rate and stirring speed in these processes. The conversion is carried out under more stringent conditions as desired, but forms a tight cake on the surface of the liquid broth. Use the cake to take some unreacted Conreno out of the broth. Example 8 The description and results of Example 8 are summarized in Table 24. Four fermentation processes were carried out, in which n-Hydroxy Conreno was produced by Conreno's biotransformation. In two of these processes (R4A and R4D), the biotransformation was operated in the same way as the processes R3 A and R3D of Example 6. In the process R4C, Conranow is generally converted to &quot; N-Hydroxy Conranow in the manner described in Example 3. In process R4B, this process is usually redundant as described in Example 4, that is, just before inoculation, the Conley and growth medium are sterilized in the fermenter, and all nitrogen and nitrogen are introduced at the beginning of the batch reaction Phosphorus is nutritional, and only a supplementary solution containing glucose is supplied to the fermenter in order to maintain the glucose content as the batch reaction proceeds. In the later process (R4B), the glucose concentration is monitored every 6 hours and the glucose solution is added as instructed to control the glucose content in the range of 0.5 to 1%. The Conrad addition schedule for these processes is listed in Table 25. -170 · This paper size applies to Chinese National Standard (CNS) A4 specification (210X297 feet) (Please read the precautions on the back before filling this page)-Order _ 593305 A7 B7 V. Description of Invention (l68) Table 24-Kang Methodology of the development experiment of Renault Biotransformation Process number R4 A R4B R4C R4D medium (g / l) Corn slurry 30 and process R4A gland: 20 and process R4A yeast extract 15 Same yeast extract: 20 Same ΝΗ4Η2Ρ〇4 3 Glucose: 2 glucose 15 〇SA: 3 ml of OSA 0.5 ml with 2.5 N NaOH pH 値 with 2.5 N NaOH Adjust to 6.5 Adjust to Conranox Sterilize Conranox and unsterilize Kangran in a fermenter Ray Blends Connor $. BI: 40 g 160 g Conorno: By bacteria in Table 25 and blended. 23.5 hours: 120 grams of schedule listed in sterilization of 40 grams 23.5 Loading hours: 120 grams of loading medium See Table 25 See Table 25 See Table 25 See Table 25 Harvest time 122 hours 122 hours 122 hours 122 hours Biotransformation 95.6% 97.6% 95.4% 96.7% (Please read the notes on the back before filling out this page} «. Printed by the Consumers' Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs -171 This paper size applies to China National Standard (CNS) A4 (210X297) Cm 593305

7 7 AB V. Description of the invention (169) Consumption Cooperatives of the Central Standards Bureau, Ministry of Economic Affairs, India, 25. When the experiment is started, the supply schedule of Conorno, glucose and growth medium is hours R4C R4A R4B R4D Conorno 200g / 2 Glucose gland and yeast-derived antibiotics 20 mg of Kang mold growth medium growth perconox / liter sterile water 50 g solution 20 g each in 1 liter of water 20 mg tetracycline 100 mg cephalosporin in 50 ml Medium (added in the form of Conreno in a slurry) See Table 24 Nutrients See Table 24 Long medium See Table 24 g / L 14 600 135 25 50 ml — — — 20 — 100--— — — 23 — __ — 120 g / 1.2 liters 120 g / 1.2 liters 26 — 100 25 — — — — 32 — 135 25 — — — ~ 38 500 120 25 50 ml — — — 44 — 100 25 — — — — 50 __ 100 25 — Sun building — — 56 — 150 25 — — — — Yi Yi 62 500 150 25 50 ml — — — 68 — 200 25 — — — — 74 — 300 25 — — — 80 — 100 25 — — — — —86 — 125 25 · — Let's go · 92 — 175 25 — — — — 98 — 150-—-— — 104 — 175 — ~ — — — 110 — 175 — — — — — 116 — 200 — — — — — -172- (Please read the notes on the back before filling out this page) Order P. This paper size is applicable to China National Standard (CNS) A4 (210X 297 mm) Printed by the Staff Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs System V. Description of the invention (no) Fermentation &amp; During the period of 4 large fermentation cycles, all are under high agitation and 2: -Γ fermented beer has become sticky within -days or after inoculation. Example 9 In Table 26, the transformation growth medium, Conrad addition plan, harvest time, and degree of transformation for performing this example are listed. Except for the following explanations, the four biotransformation processes were generally performed by the method described in the process r4b of Example 8. In process R5B, a thirsty roulette propeller is used for agitation. In other processes, a downwardly drawn seawater propeller is used instead. The downward pumping action causes the broth to flow axially into the center of the fermenter, reducing cake formation. In process r 5 D, methanol (200 ml) was added immediately after seeding. Because the Connor in the fermenter is sterile, all nutrients except glucose are added at the beginning of the batch, eliminating the need for a continuous supply of nitrogen, phosphorus, or antibiotics. -173- This paper size applies to China National Standard (CNS) A4 (210X297 mm) (Please read the precautions on the back before filling this page}

593305 A7 B7 Consumption cooperation with employees of the Central Bureau of Standards of the Ministry of Economic Affairs. Printed 5. Description of the invention (171 Table 26-Method for carrying out the experiment on the biotransformation on a scale of 10 liters. Process description R5 A R5B R5C R5D culture medium Per litre) corn pulp yeast extract ΝΗ4Η2Ρ〇4 glucose OSA pH 値 30 15 3 15 0.5 ml adjusted with 6.5 N NaOH to 6.5 Same as the process R5A gland: 20 yeast extract: 2 glucose: 20 〇SA: 3 The liter was adjusted to 2.5 with 2.5 N NaOH. It was the same as the process R5A. Loaded into Conranos. Sterilized 160 grams of Conranos in the fermenter. Sterilized 160 grams of Conranos in the fermenter. Sterilized 160 grams of Conranos in the fermentation. 160 grams of Conreno sterilization medium supply glucose supply glucose supply glucose supply glucose supply glucose harvest time 119.5 hours 119.5 hours 106 hours 119.5 hours Biotransformation 96% 94.1% 88.5% 92.4% In order to maintain the infiltration of the solid phase growing on the liquid surface 9 to 6 hours after the start of the batch, add growth medium (2 liters) to each fermenter. By adding the growth medium Using a downwardly drawn thruster (process r 5 B), the brigade cannot completely overcome the mixing problem, but the results of the process confirm the feasibility and advantages of the method and show that it can provide satisfactory mixing according to traditional practices- 174- The size of this paper applies to Chinese National Standard (CNS) A4 (210X29? Mm) (Please read the precautions on the back before filling this page)

1T 593305 A7 B7 V. Description of the Invention (I72) Example 1 0 Three biotransformation processes were carried out substantially in the manner described in Example 9. List the transformation medium, Conranos addition schedule, harvest time, and the degree of transformation of the process in this example in Table 27. Table 27. Description of the Experimental Methods for Biotransformation on a 10-liter Scale Process Number R6A R6B R6C Medium (g Per litre) Corn slurry 30 Same as R6A gland: 20 yeast extract 15 same yeast extract: 2〇NH4H2P〇4 3 glucose: 20 glucose 15 OSA: 0.5 ml OSA 0.5 ml with 2.5 N NaOH adjusted to pH 値 with 2.5 N NaOH adjusted to 6.5 6.5 Load Conley in the fermenter will be 160 in the fermenter will be 160 in the fermentor will be 160 grams of Concord sterilized Kronor sterilized Reynolds sterilized medium supply glucose: supply glucose at 71 ; Supply glucose; no additional hours 1.3 liters medium 96 hours 0.5 liters culture addition and 0.8 liters of sterile water-based and 0.5 liters of sterile water: harvest time 120 hours 120 hours 120 hours biotransformation 95% 96% 90% material balance 59 % 54% 80% Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) In process R6A, After 71 h addition of growth medium (1.3 L) and sterile water (0.8 l), in order to submerge mycelial cake on the surface of the liquid broth growth. For the same purpose, in the process R6 B, add growth after 175 hours -175- This paper size applies Chinese National Standard (CNS) A4 specifications (210X 297 mm 1 593305 V. Description of the invention (Π3) Medium (0.5 Liters) and sterile water (G5 liters). The material balance data show that reducing the cakes built on the liquid surface can determine a better material balance. Example 1 1 A fermentation process was performed to compare Conranow's pre-sterilization and conversion Conreno is attenuated with the growth medium in the fermenter. In process R7A, as illustrated in Figure 2, under conditions comparable to those of process r2C, R2D, R3 A, R3B, R3D, R4 A, and R4D This process is completed. Process R7B is completed under the conditions comparable to those of Examples 4, 9, and 丨, and Process R4B, as illustrated in Figure 3. The undesired transformational growth of this example is listed in Tables 28. Culture medium, Conley language addition plan, harvest time and degree of transformation: (Please read the notes on the back before filling out this page)-Ordered by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs to print 176- ------ This paper size Applicable in China Standard (CNS) A4 (210X297 mm) 593305 V. Description of the invention (174) Table 2 Experimental method description of biotransformation on a scale of 10 liters-Process number R7A R7B medium (g / L) Corn slurry 30 and Process R7 A same yeast extract 15 NH4H2P〇4 3 glucose 15 OSA 0.5 ml pH 値 adjusted to 2.5 NNaOH to 6.5 Load Conranox outside the fermenter 160 g of Conrad sterilized and blended in the fermenter Hano sterilization medium supply and supply of glucose; Conreno with supply of glucose: 1.6 liters of growth medium with or without addition to harvest time 118.5 hours 118.5 hours Biotransformation 93% 89% (Please read the precautions on the back before filling out this page The Ministry of Central Standards Bureau ’s Consumer Cooperative printed a material balance of 89.5% based on the final sample obtained from process R7 B, showing no significant loss or degradation of enzymes during biotransformation. Two processes were identified The mixing is appropriate. During the first 80 hours, the residual glucose concentration is within the desired control range of 5 to 10 gp 1 Above. The efficiency of the process is obviously not affected by the muffins stacked on the liquid surface of the two fermenters. Example 1 2 -177- This paper size applies to China National Standard (CNS) A4 (210X297 mm) 593305 Ministry of Economic Affairs Printed by the Consumer Standards Cooperative of the Central Bureau of Standards 5. Description of the Invention (175) According to the summary in Table 29, a series of 1 liter extraction processes are used to determine the extraction efficiency. In each process, ethyl acetate (1 L / L fermentation volume) was used to extract steroids from the mycelium. Each process is continuous and extracted twice. Based on RP-HPLC, approximately 80% of the total steroids were recovered during the first extraction; and the recovery was increased to 95% by the second extraction. The third extraction will recover the other 3% steroids. The remaining 2% is lost in the supernatant liquid phase. Extract with suction to dryness, but rinse without any additional solvents. If for method economic reasons, solvent hunting will improve the recovery from the original extraction. Table 29-Recovery of 11-Hydroxy Conreno in 1 liter extraction (% of total) Process number First extraction Second extraction Third extraction supernatant R5A 79% 16% 2% 2% R5A 84 % 12% 2% 2% R4A 72% 20% 4% 4% R4A 79% 14% 2% 5% R4B 76% 19% 4% 1% R4B 79% 16% 3% 2% R4B 82% 15% 2% 1% Average 79% 16% 3% 2% Methyl isobutyl ketone (MIBK) and toluene were evaluated on a 1 liter broth scale as the extraction / crystallization solvent for 1 1 α-hydroxyconoreno. Using the extraction scheme as described above, the extraction efficiency and crystallization efficiency of MIBK and toluene and ethyl acetate were compared. -178- This paper size applies to China National Standard (CNS) A4 (210X 297mm) (Please read the notes on the back before filling out this page, 1T-593305 A7 B7 Printed by the Staff Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 2. Description of the Invention (I76) As part of the method for evaluating Figures 2 and 3, the study of particle size was carried out on the Conreno enzymes provided at the beginning of the fermentation cycle of each method. Prior to introducing Conranow into the fermentor, the Conranow, which supplies the method of Figure 1, was micronized. In the process, Conranow was not 'subtracted' by adding antibiotics to control the growth of unwanted microorganisms. Figure 2 and In the process of Figure 3, Conreno was sterilized at the end of the reaction. In the process of Figure 2, the sterilization was completed in the blender before introduction into the fermenter. In the process of Figure 3, at the beginning of the batch reaction, The fermentor sterilizes the suspension of Conorno in the growth medium. As discussed earlier, sterilization can easily cause the accumulation of Conreno particles. Because of the limited solubility of Conreno in aqueous growth media, The productivity of this method depends on the transfer of clumps from the solid phase, and it can therefore be expected to depend on the interfacial area provided by the solid particulate enzymatic substance, which depends on the particle size distribution. These considerations became graphs from the beginning Obstacles to the methods 2 and 3. However, the agitation effect in the blender of Fig. 2 and the fermenter of Fig. 3 and the effect of the shear pump used for migration in Fig. 2 were found to make the particle size The range is quite close to the agglomeration degradation of the non-sterilized and micronized Conorno supplied by the method of Figure 1. By these three processes, the beginning of each reaction cycle can be shown in Table 30. Three; f, the same Particle distribution vf of Conranos sample, 1 strict | 1 »1 1 V specimen 45-125 microns &lt; 180 microns average size micron process number # biotransformation% Conranow loading 1 75% 95% — R3C: 93.1 % (120 hours) -179- This paper size applies to Chinese National Standard (CNS) A4 (21〇χ 297mm) (Please read the precautions on the back before filling out this page} 1 «-Order · 593305 A7 B7 5 Description of the invention (177) R4C: 96.3% (120 hours) % 77.2% 139.5 β3Α ·· 94 · 6% (120 hours) R3B: 95.2% (120 hours) Sterilized d sample 24.7% 65.1% 157.4 R4B: 97.6% (120 hours) R5B: 93.8% (120 hours) ( Please read the precautions on the back before filling in this page} From the data in Table 30, you will notice that the agitator and shear pump can effectively reduce the average particle size of the sterilized Conorno The size of the enzymes is the same, but the remaining significant size differences will benefit the unsterilized enzymes. Despite these differences, reaction efficiency data show that pre-sterilization methods are at least as productive as the method in Figure 1. In the method of Figure 2, further benefits can be obtained by some further reduction and control of particle size steps, such as wet milling of sterilized Conley #, and / or by pasteurization rather than sterilization. Example 14 A seed culture was prepared in the manner described in Example 5. After 20 hours, the mycelium in the inoculated fermenter was soft, with 40% ρ μV. Its ρ Η 値 is 5 · 4 and unused 1 4 · 8 gp 1 glucose is left. Printed by a Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs A transforming growth medium (35 liters) having the composition shown in Table 20 was prepared. In the preparation of the supply medium, the glucose and the yeast extract were respectively sterilized and mixed into one portion at a starting concentration of 30% by weight of glucose and 10% by weight of the yeast extract. 1) 11 値 of this supply was adjusted to 57. Using this medium (Table 20), two biotransformation processes were performed to convert Conranow to 1 1 -hydroxyl Conax. Each process is equipped with a stirrer, including a Rushton full-wheel propeller and two Lightnin, A31q ^ feeders. 180- This paper size applies to China National Standard (CNS) A4 specifications (21 〇X 297 male thin ) 593305 A7 _____ B7 V. Description of the Invention (l78) Operating in a 60-liter fermenter. The growth medium initially loaded into the fermenter was 35 liters. Add micronized but non-sterile Conranow to a starting concentration of 0.5%. The seed culture prepared by the method described in Example 5 was used to inoculate the culture medium in the fermenter at an initial inoculation ratio of 2.5%. Fermentation is performed at a temperature of 28 ° C, agitation of 200 to 500 rpm: agitation rate &apos; 0.5 vvm aeration rate, and a back pressure sufficient to maintain at least 20% by volume of dissolved oxygen. The transformation medium developed during the production process is in the form of very small rounded spheres (approximately 1-2 mm). Conreno and supplementary nutrients were continuously supplied to the fermenter in the manner described in Example 1. Nutrition was added every four hours at a ratio of 3.4 grams of glucose and 0.6 grams of yeast extract per liter of broth in the fermenter. In Table 31, the main aeration rate, stirring rate, dissolved oxygen amount, PMV, and pH 値 are set during the period of each process in this example, and the glucose is added during the batch reaction. . Table 3 2 shows the appearance of Conley's transdermal effect. The process r 1 1 a was aborted after 46 hours; the process r 1 1 B continued to 96 hours. In the later process, a transition of 930/0 was reached at 81 hours; the supply was added again at 84 hours; then the supply was discontinued. Note that a noticeable change in viscosity occurred between the time the supply was stopped and the end of the process. (Please read the precautions on the back before filling this page) 1 «· Ordered by the Central Consumers Bureau of the Ministry of Economic Affairs and printed by the Consumer Cooperatives -181- This paper size applies to China National Standard (CNS) specifications (210X ^ 97 mm) ~~ 593305 A7 B7, Invention Description (Π9) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 3 1 _ H Rong R 1 1 A Time Air (1pm) rpm% DO Back Pressure PMV (%) pH 値 Glucose (g Per liter) 0.1 20 200 93 0 2 6.17 5.8 7 20 200 85.1 0 5 6.03 5.5 12.4 20 300 50.2 0 5.43 21.8 20 400 25.5 0 38 6.98 0 29 20 500 17 0 35 5.22 30.2 20 500 18.8 10 5.01 31 20 500 79 10 4.81 1 35.7 20 500 100 10 45 5.57 0 46.2 20 500 23 6 45 5.8 1 Total glucose: 27.5 g / L total yeast extract: 8.75 g / L Fermentation R11B Time Air (1pm) rpm% DO Back pressure PMV ( %) pH 値 glucose (g / l) 0.1 20 200 92.9 0 2 5.98 5.5 7 20 200 82.3 0 5 5.9 5 12.4 20 300 49.5 0 5.48 21.8 20 400 18 0 40 7.12 0 29 20 500 36.8 0 35 5.1 3 35.7 20 500 94.5 10 4.74 0 46.2 20 500 14.5 6 45 5.32 2 55 20 500 16.7 10 5.31 0.5 58.6 20 500 19.4 15 5.32 1 61.9 20 500 13 15 40 5.36 2 71.7 20 500 13 15 42 5.37 0 81.1 20 500 22.9 15 5.42 2.5 85.6 20 500 22 15 45 5.48 1 97.5 20 500 108 15 45 6.47 0 117.7 20 500 15 7.37 0 Total glucose: 63 g / L Total Yeast Extract: 14.5 g / L -182- This paper size applies Chinese National Standard (CNS) A4 specification (210 × 297 mm) (Please read the precautions on the back before filling in this Page 丨 «, 1T 593305 V. Description of the invention (180) Printed by the Consumers' Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 32 Fermentation process R11A: Conranow conversion concentration (g / L) Conversion calculation Per liter / hour) Sample time 〇Η-Cono Reno Cono Reynolds (%) (g / L) Calculated 値 Measured R11A-0 0.10 0.00 5.41 5.41 R11A-7 7.00 0.18 4.89 5.07 3.58 0.18 0.03 0.03 R11A-22 21.8 2.02 2.12 4.14 48.75 2.44 0.15 0.12 R11A-29 29.00 3.67 4.14 7.81 47.03 4.48 0.28 0.23 R11A-36 35.7 6.68 1.44 8.12 82.27 7.74 0.49 0.45 R11A-46 46.20 7.09 0.41 7.51 94.48 8.59 0.08 0.04 Fermentation process R1 1B : Conreno conversion effect concentration (g / L) Conversion calculation ΗΟ-Conoreno conversion rate (g / L / hour) Sample time 〇Η-Conoreno Conreno total (%) (g / L) Calculation 値 Measurement 値 R11B- 0 0.10 0.00 5.60 5.60 R11B-7 7.00 0.20 4.98 5.18 3.78 0.19 0.03 0.03 R11B-22 21.80 2.51 2.46 4.97 50.49 2.52 0.16 0.16 R11B-29 29.00 4.48 16.99 21.47 20.87 4.69 0.30 0.27 R11B-36 35.70 8.18 10.35 18.53 44.16 9.70 0.75 0.55 R11B -55 55.0 17.03 13.20 30.23 56.33 19.50 0.32 0.36 R11B-59 58.6 20.80 11.73 32.53 63.95 21.97 0.69 1.05 R11B-62 61.9 22.19 8.62 30.81 72.02 24.50 0.77 0.42 R11B-72 71.7 26.62 3.61 30.23 88.06 29.46 0.51 0.45 R11B13-81 92.97 30.32 0.09 0.05 R11B-86 85.6 26.87 2.02 28.88 93.02 30.11 -0.04 -0.06 R11B-97 97.5 23.95 1.71 25.66 93.34 30.22 0.01 -0.25 R11B-118 117.7 24.10 1.68 25.79 93.47 30.26 0.00 0.01 (Please read the notes on the back and fill in (This page)

、 1T -183- This paper size is applicable to Chinese National Standard (CNS) A4 specification (210 × 297 mm) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 593305 A7 Γ ________ Β7 V. Description of the invention (181) iM ± 5_ According to the above The general method of testing the biotransformation efficiency of various cultures to convert Conranow to 11α-Conranow. Separate working cell banks of AspeH1hl, niger ATCC 1 1394 and Rhizopus arrhizus ^ ATrr 11145 and (JB ^ hizopus stolonifer) ACTT 6227b were prepared by the method described in Example 5. A spore suspension (1 ml) from a working cell bank was used to inoculate a growth medium (50 ml) having the composition listed in Table 18 and placed in an incubator. By fermentation at 26 ° C for approximately 20 hours, Prepare the seed culture in an incubator. Stir the incubator at 200 rpm. Use an aliquot (2 ml) of the seed culture of each microorganism to inoculate a transformation containing the growth medium (30 ml) of Table 18 Flasks. Use each culture to inoculate two flasks for a total of six. Dissolve Conreno (200 g) in methanol (4 ml) at 36 t, and aliquot 0.5 ml of this The solution was introduced into each flask. The biotransformation was normally completed under the conditions described in Example 5, and a 50% by weight glucose solution (1 ml) was added daily. After the first 72 hours, the bacteria in the individual transformation fermentation flasks were added. The following observations were made on the development of the body: ATCC 11394-Good and balanced growth ATCC 11145 _ Good growth in the first 48 hours, but mycelial clumps formed spheres and no significant growth in the next 24 hours. ATCC 6227b-Good Growth; Mycelium clumps form spheres. Take a sample of broth and analyze the degree of biotransformation. After three days, the fermentation of ATCC 1 1 3 94 was used to provide 80 to 90%! "-Jiji-184- This paper size applies Chinese National Standard (CNS) grid (210X 297mm) '~ (Please read the precautions on the back before filling this page), \ 呑 ^ 1. 593305

AB V. Description of the invention (182) Connor's transformation; ATCC 1 1 145 provides 50% of the transformation; ATCC 6 2 2 7 b is for 80 to J 9 0 〇 / 〇 spoon transfer &gt; ί dagger. Example 16 Using the method described in Example 15 in general, other microorganisms were tested for their efficiency in converting Conranow to 1 1 α-hydroxyconranol. The organisms tested and the test results are shown in Table 33: Table 33-Cultures subjected to bioconversion tests to convert Conranos to 11-heart light Conranos ATCC numbered medium 1 Results approximately conversion rate 1145 CSL + 50% Saccharomyces cerevisiae 6227b CSL + 80-90%-S. licheniformis 11267 CSL + 50% 80% Black fungus 11394 CSL + 80-90% Pseudomonas flavus NRRL405 CSL + 90% 18500 CSL + 90% Bacillus subtilis 31028 P &amp; CSL 0% 0% Bacillus subtilis 31028 CSL-0% 0% Bacillus sp. 31029 P &amp; CSL-0% 0% Bacillus 31029 CSL 0% Bacillus megaterium 14945 P & SCL + 5% 80% * Macrobacterium 14945 CSL + 5% 10% * Monophyllum pink 12519 CSL + 80% * 90% * Monophyllum pink 8685 CSL + 80% * 90% * Streptomyces freundii 10745 CSL + &lt; 5% &lt; 10% Streptomyces freundii 10745 TSB-氺 * -185- This paper size applies to China National Standard (CNS) A4 specifications (2丨 〇 &gt; &lt; 297 mm) (Read the precautions on the back before filling in this page)-、 1Τ Staff of the Central Standards Bureau of the Ministry of Economic Affairs Cooperatives printed 593,305

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7 B V. Description of the invention (183) Streptomyces lilacinus 13664 CSL-0% * Streptomyces lilacinus 13664 TSB-0% 0% Nocardiodes simple 6946 BP-0% 0% simple Soil Mycelia 13260 BP * * Pseudomonas 14696 BP-Pseudomonas 14696 CSL + &lt; 5% &lt; 10% Pseudomonas 14696 TSB-0% * Pseudomonas 13261 BP + 10 10% Pseudomonas kusifa 13262 BP # &lt; 10% Pseudomonas putida * forms other unidentifiable products 15175 BP 0% 0% (please read the first Note: Please fill in this page again. 1 Medium: CSL-corn slurry; TSB-trypsin-digested soy broth; P & CSL-gland and corn slurry; BP · beef extract and gland. Consumption cooperatives tested various microorganisms to convert Conley to 9 α-Kyrelino. The fermentation medium for this example was prepared as listed in Table 34: Table 34 Soy meal: Dextrose 20 Grams of soybean meal, 5 grams of NaCl, 5 grams of -186- This paper size applies to Chinese national standards (CN S) A4 specification (210X297 mm) 593305 A7 B7 -------------— ~~ -——— 5. Description of the invention (184) Yeasts printed by employees' cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs Extract 5 g KH2P〇4 5 g water to 1 liter pH 値 7.0 Gland / yeast extract / glucose: 40 g glucose bactopeptone 10 g yeast extract 5 g water to 1 liter Mueller-Hidden (Mueller -Hinton): 300 grams of beef infusion casein amino acid "· 5 grams of starch 1.5 grams of water to 1 liter of genus Pseudomonas grow on soybean meal meal medium and glandular-yeast extract · glucose And Genus Bacteroides are grown in soybean meal (for biotransformation, 0.9% by weight sodium formate) and Mueller-Hidden broth. Initial cultures were inoculated with frozen spore mother liquor (at 25 20 ml of soybean meal in a 0 ml Erlenmeyer flask). Cover the flask with milk filter paper as a biological protection. Inoculate the metabolic culture (also in a 250¾L Erlenmeyer flask with the starting culture (24 or 48 hours long)) 20 ml)-with a fork volume of 15%-in the plus The reaction for conversion of the steroid by the use of quality prior to the enzyme, the culture which is 24 to 48 hours. Dissolve / suspend in Reynolds in methanol (20 mg / ml), filter sterilize -187- This paper size applies to China National Standard (CNS) A4 specifications (210X297 Gonglu Γ 593305 Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economy A7 -—_____ B7__ 5. Description of the invention (185) ~~, and added to the culture to a final concentration of 0.1 · g / ml. All roasts are in a controlled greenhouse at 2 6 π At 60%, shake at 2500rpm (2 &quot; throwing). Biotransformation is harvested 5 to 48 hours after adding the substrate, or 24 hours. By ethyl acetate (2 3 ml ) Or dichloromethane is added to the fermentation flask to start the harvest. Then shake the flask for two minutes and pour the inner grain of each flask into a 50 ml conical test tube. To separate the phases, centrifuge at 4000 rpm at room temperature Test tube for 20 minutes. Move the organic layer from each test tube to a 20 ml borosilicate glass vial and evaporate it in the air with variable speed. Cap the vial and store at -20 ° C. To obtain a substance for structure determination By adding a shake flask The number of effects was increased to 25, and the biotransformation effect was scaled up to 500 ml. At the time of harvest (24 or 48 hours after the enzyme was added), ethyl acetate was added to each flask and the flask was Cap and return to shaker for 20 minutes. Then pour the contents of the flask into a polypropylene bottle and centrifuge to separate the phases, or into a separatory funnel that allows the phases to be separated by gravity. Place the organic phase Dehydration produces a crude steroid extract contained in the reaction mixture. The reaction products are first analyzed by thin-layer chromatography on a silica gel (25 micron) on the back fluorescent plate (254 nm). Ethyl acetate (500 μl) was added to each vial of dehydrated ethyl acetate extract of the reaction mixture. Further analysis was performed by high-efficiency liquid chromatography and mass spectrometry. At 9 5: 5 vol / Chromatographic plates are developed in a volume of gas imitation / methanol medium. Further analysis is carried out by high-efficiency liquid chromatography and mass spectrometry -188- This paper is in accordance with China National Standard (CNS) A4 specifications (2 10 &gt; &lt; 297 mm) (Please read the precautions on the back before filling this page) 丨 ^ »· -ding,,--1. I 1 II --- 593305 Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs Preparation A7 B7 V. Description of the invention (186). Waters HPLC with Millennium software, photodiode array detector and automatic sampler. Reverse phase HPLC using Waters NovaPak C -1 8 (4 micron particle size) Radi alPak 4 mm centrifuge tubes. The initial 25-minute linear solvent gradient in the column was water: acetonitrile (7 5: 2 5), and at the end it was water: acetonitrile (2 5: 7 5). Then a three minute gradient to 100% acetonitrile and a 4 minute isocratic rinse before updating the column in the starting conditions. For LC / MS, ammonium acetate was added to the acetonitrile and water phases at a concentration of 2 nM. Chromatography is not significantly affected. The eluate from the column was split into 22: 1 and most of the material was sent to the ρOA detector. The remaining 4.5% was sent to the electron spray ionization chamber of the Sciex API III mass spectrometer. Mass spectrometry analysis was performed in positive mode. For common analysis of UV and MS data, a similar data line was transferred from the PDA detector to the mass spectrometer in a single wavelength chromatogram on HPLC. Mass spectrometry was used to classify hydroxylated enzymes. Two prospective acylated Conreno, 11-hydroxyl and 9α-hydroxy-Conreno, lose water in a consistent manner but at different frequencies and can be used as diagnostics. Furthermore, 9α-hydroxyconoreno is more likely to form an ammonium addition compound than ιαα-hydroxyconreno. Listed in Table 35 is a summary of TLC, HPLC / UV, and LC / MS data from Conranox fermentations, showing that the tested microorganisms were effective in converting Conranow to 9α-hydroxy Conranow. The better microorganism is Corvnespora cassiicola ATCC 16718. -189- This paper size is in accordance with China National Standard (CNS) A4 specification (2〗 〇297 × 297 mm) (Please read the precautions on the back before filling this page), ^ τ 593305 B7 V. Description of Invention (Μ?) Ministry of Economy Printed by the Central Standards Bureau Consumer Cooperatives Table 3 5-Summary of TLC, HPLC / UV, and LC / MS data on Conranox fermentation 9 Based on the evidence of OH-Conradano Cultures TLC ink at 9 Han OH-AD HPLC at 9 a OH-Conoreno_ Peak w / UV MS: 357 (M + H), 339 (-H20) &amp; 375 (+ NH4) Pleurotus candida ATCC6647 η yy / n Gray-green pear head Mold ATCC 22752 η Mucor elegans ATCC 6476 tr y tr Pseudomonas aeruginosa ATCC 1030 tr A. fumigatus ATCC 26934 tr yn Puccinia striiformis ATCC 11267 tr yy Black fungus ATCC 16888 nyy Black fungus ATCC 26693 nyn brown yellow Pseudomonas ATCC 18500 nyn Snail Curl Shell ATCC 10195 tr tr y / n Testosterone clinomonas tr tr n -190- (Please read the notes on the back before filling out this page) This paper size applies to China National Standard (CNS) A4 (210X297 mm) 593305

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7 B V. Description of the invention (188) Printed by fComomona testosteroni) (Searle) ATCC 11996 Corynespora mandshurica ATCC 16718 yyy Brachytium eryngii ATCC yyy 8688a ATCC yyy 3655 Elegant gram silver bacterium ATCC yyy 9245 Curvularia sp. ATCC 22921 nyy / n Curvularia sp. ATCC 12071 ynn Searle tr nn ATCC 11011 Epidermococcus hermicella ATCC yyy 12722 HEpicoccum tr tr tr orvzae) ATCC 12724 Fusarium oxysporum ATCC 7601 tr ) ATCC 10097 Tococcus ovum ATCC 22822 y yUV? Y Chlorella parasite IMI 109891 yyy Lipomvces n lipofer, ATCC 10792 iMelanospora omata tr nn (Please read the precautions on the back first (Fill in this page) -191- This paper size applies to China National Standard (CNS) A4 (210X 297mm) ) 593305 5. Description of invention (189) Employees' cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs of the People's Republic of China. ATCC 26180 Mortierella aureus ATCC 42613 yyn White mold ATCC 1027a η White mold ATCC 4605 tr yy Mycobacterium sporum ATCC 6842 Warts P. aeruginosa ATCC 9095 tr tr y Searle n tr n ATCC 12674 Searle yyn Agrobacterium coral ATCC 19070 n Pseudomonas aeruginosa ATCC 46579 nyn Penicillium chrysogenum ATCC 9480 n Penicillium sp.ATCC 24550 yyy / n Penicillium purpuratum ATCC 46581 tr yy Aso-Oerlikhiomyces ATCC tr y tr 6651 Mycelium canine ATCC 26150 n tr tr Brachyspira (Thvcomvces yyy / n blakesleeanus, Mycobacterium ATCC 12231 yyy / n Rhizopogonus. Rhizococcus sp. ATCC 11145 tr yn Rhizoctonia sp. ATCC 6227b n E.coccus ATCC 14887 n tr n E.coccus ATCC 21329 tr tr n (Please read the note on the back first Please fill in this page for more details) -192- This paper size is applicable to China National Standard (CNS) A4 specification (210X297mm) 593305 V. Description of invention (19〇) Economy Printed by the Consumer Standards Cooperative of the Central Bureau of Standards of China Affinis Septorto Mesa η yUV? Y / n iSeptomvxa affinis) ATCC 6737 Two-color staphylococcus ATCC 12672 yyy / n Streptomyces californicus ATCC 15436 η% Streptomyces senecio fStreptomvces cinereocrocatus) ATCC 3443 Streptomyces azure ATCC 10147 η Streptomyces striatus ATCC 25453 Streptomyces freundii ATCC 10745 η Grey key mold psychrophile η fStreptomvces griseus subsp. Eriseus) ATCC 13968 Gray Streptomyces ATCC 11984 η Streptomyces hairui (Please read the notes on the back before filling in this page. Order ^ 1. -193-This paper size applies to Chinese National Standard (CNS) A4 specification (210X297 mm) 593305 kl B7 V. Description of the invention (191) fStreptomvces hvdro genans ) ATCC 19631 Streptococcus hygroscopicus ATCC 27438 yyy Streptomyces lilacinus Panlab 105 η oligospore chain Mold ATCC 25489 η Streptomyces spp. ATCC 25489 η tr tr Streptomyces roseus ATCC 13400 Streptomyces magnificentus ATCC 27465 η Stvsanus microsporus ATCC 2833 Conidial cephalosporin ATCC 18192 η Thamnidium elegans) ATCC 18191 Tyrosoma pyriformis ATCC 8992 y tr y Clostridium sp. ATCC 13807 n Trichoderma viride ^ ATCC 26802 η Monomonospora pink ATCC 12543 tr yy / n Cocoa verticillium ATCC 12474 y tr tr Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) Example 1 8 According to the general method described above, test various cultures to bioconvert androstenedione to 1 1 α- # 垔 基 andosterone dione potency.

Basically, the method described in Example 4 was used to prepare Pseudomonas flavus NRRL-194- This paper size applies the Chinese National Standard (CNS) A4 specification (210 × 297 mm 1 593305 A7 _________B7 V. Description of the invention (192) 405 ( (ATCC 1 8 5 00); black fungus ATCC 1 1 394; small nest fungus ATCC 1 1 267; wine bacterium fungus ATCC 11145; Working cell bank. A spore suspension (丨 ml) obtained from the working cell bank was used to inoculate a growth medium (50 ml) having the composition listed in Table 18 and placed in a 7-dish phase. Warm-phase spermatozoons ferment for about 20 hours at 26 C to prepare seed cultures. Stir the incubator at 200 rpm. Use an aliquot (2 ml) of each seed culture of each microorganism to Inoculate a transformation flask containing the growth medium (30 ml) of Table 15. Inoculate two flasks with each culture, for a total of 16. Dissolve androgens dione 300 g at 36 ° C) in Methanol (6 ml), and the solution was 0.5 ml Parts introduced into each flask. The biotransformation is usually carried out under the conditions described in Example 6 for 48 hours. A sample of the broth was collected after 48 hours and extracted with ethyl acetate as in Example 17. Ethyl acetate was concentrated by evaporation, and the sample was analyzed by thin-layer chromatography to determine whether a layer with a similar standard of 1 1 α _ few androstenedione dione (Sigma Chemical Co., St. Louis) appeared. Analytical product of fullness. The results are shown in Table 36. M + &quot; is a positive fruit. Table 36: Male and female associations-one-to-one to 1 1 α-total androgenous one-to-one biotransformation culture ATCC numbering medium TLC results Saccharomyces cerevisiae 11145 CSL Saccharomyces cerevisiae 6227b CSL + small nested tadpole Bacteria 11267 CSL + -195- Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the precautions on the back before filling out this page) This paper size applies to China National Standard (CNS) A4 (210 '乂 297 mm) 593305 A7 B7 V. Description of the invention (193 Black fungus 11394 CSL + Pseudomonas flavus NRRL 405 CSL + Pseudomonas flavus 1 8500 CSL + Pink monospore 125 19 CSL + Pink monospore 86 85 CSL + ketene A compound with Rf 値 like the standard of 11-hydroxyandrostadione was produced in the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs of the People's Republic of China. Using the same procedure described above, the test was carried out to expose the fungus NRRL 40.5 (ATCC 18500), and the culture product was separated, purified by normal-phase silica gel column chromatography, using methanol as a solvent, and analyzed by thin layer chromatography. The TLC plate was Whatman K6F Silicone 60 Angstrom, 1 (^ 20 size, 250 micron Thickness. The solvent system was methanol: aerosol, parent 95 vol / vol. Analysis of the crystalline product and n-hydroxy ^ complexedione standard by LC-MS and NMR spectrometer. Both compounds gave the same appearance and molecular weight Example 19 The various microorganisms were tested for their effectiveness in converting Mexrenault to old. Ricci Mexrenault. The fermentation medium of this example was prepared as described in Table 34. Fermentation conditions and analysis methods were the same as in Example 17. The same. The tlc plate and the solvent system are the same as described in Example 18. The principle of the chromatographic analysis is as follows: iH minus Reynolds and U ^ Conconnol have the same chromatographic fullness. «&Lt; &amp; 广- Ji Kang Tian Nuo and 9 ^ Li Ji Kang Renuo show the same full-scale mode as 11 "-Kiji Tian Tian 'hexadione and u factory hydroxyandrostadione. Therefore η (Please read the precautions on the back before Fill out this page) Order ^ 1. M's Zhang scale suitable financial Guanjia -196- 593305 7 Β V. Description of the invention (194) Employees' Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs Consumer Cooperative India-Hydroxy Max Reynolds should have something like 9 α-Hydroxy Conreno has the same mobility. Therefore, the The compounds extracted from the growth medium were analyzed for 9α-hydroxyconoxeno as a standard. The results are shown in Table 37. Table 37 Summary of data for the formation of 11 /?-Hydroxymaxox Reynolds from Max Reynolds Microbial culture medium 1 Spot characteristics 2 P. aeruginosa ATCC6647 M, S strong black fungus ATCC 16888 S, P weak (S)? (P) silkworm chlamydia ATCC7159 P strong silk fungus ATCC 13144 s, p?,? Staphylococcus aureus IMI 038560 Weak short spiny Pleurotus eryngii ATCC 8688a S, p strong Escherichia sp. Microemulsion ATCC 3655 S, p strong elegant Pleurotus eryngii ATCC 9245 s, p. Lunatatus ATCC 12017 s T. oviparus ATCC 22822 s, p. Penicillium ATCC 24550 S, p. Purpureus ATCC 46581 s, p. Aso-Ourlikii mycobacterium IFO 6651 s, p. Marcescens ATCC 14887 M Saccharomyces faecalis ATCC 11635 M, SF Streptomyces faecalis ATCC 27438 M, SF Streptomyces spp. ATCC 25489 M? SF Mycobacterium sp. ATCC 18191 S, P ATCC 8992 S, P weak pink monotetra ATCC 12543 P, S weak (p) ,? (s) (Please read the notes on the back before filling out this page) -197- This paper size applies to China National Standard (CNS) A4 (210X297 mm) 593305 A7

Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs f = Mueller-Hidden P = PYG (gland / yeast extract / glucose) s = soybean meal s F == soybean meal and formate 2? = Without acceptance The difference in the enzymatic control group is uncertain. These data suggest that most of the organisms listed in this table can produce products similar to or the same as 1 1 々-hydroxymaxex Renault. tAH 2 〇 Plan 1: Step 2: Preparation of 5, R (5, from), 7, /?-20, _aminohexadecyl-U, /?-hydroxy_10'a, 13, α_dimethyl 3,5,5-dioxo [furan-2 (3h), 17, (511 ')-[7,4] methylenefluorene [411] cyclopenta [&amp;] phenanthrene] -5, -nitrile. A 50-gallon glass-lined reactor was charged with 61 _2 liters (578 kg) of DMF, followed by 23.5 kg of 11-hydroxyconoreno 1 and stirred. To the mixture was added 7.1 kg of lithium vaporized, and the mixture was stirred for 20 minutes, charged with 16.9 g of acetone cyanohydrin, and then charged with 5.1 kg of triethylamine. The mixture was heated to 85 ° C and maintained at this temperature for 1 3 to 1 8 hours. After the reaction, 3 5 3 liters of water was added, followed by 5.6 kg of sodium bicarbonate. The mixture was cooled to 0 ° C, transferred to a 200-gallon glass-lined reactor, and stopped slowly with 130 kg of a 6.7% sodium hypoxia solution. The product was filtered and rinsed with 3x40 liter portions of water. 21.4 kg of product amidine were obtained. -198- This paper size applies to Chinese National Standard (CNS) A4 (210X 297 mm) f Please read the notes on the back before filling out this page)-Order · 593305 A7 B7 V. Invention Description (196

LICK DMF, EI3N, Acetochlorohydrin β5 0, 8-15 h Step 1

K

Example 2 1 Scheme 1: Step 2: Preparation of 4, S (4 '^ 〇, 7' from -hexadecane-11 'from -hydroxy-10 · point, 13 * 々 · dimethyl-3', 5, 20'-Dioxo [furan-2 (3H), 17 * /?-[4,7] methylpyrene- [17H] cyclopenta [a] phenanthrene] · 5 * /? (2'Η) _ nitrile A 200 gallon glass-lined reactor was charged with 50 kg of enamine 2, approximately 445 liters of 0.8 N dilute hydrogen acid and 75 liters of methanol. The mixture was heated to 80 ° C for 5 hours and cooled to 0 ° C. 2 hours. The solid product was filtered to obtain 3 6.5 kg of anhydrous dione. 〇Q (Please read the precautions on the back before filling this page) Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

HCIf QH3〇Hr HaO »OeCr 5h ____________1 Step 2

Scheme 1: Step 3 A: Preparation of 1 1 a, 1 7 Jiang-dihydroxy_ 3 _oxoprogestin-4 -en-7 pan, 2 1 _ hydrogen dicarboxylic acid methyl ester, r-lactone 0 -199- This paper size applies to Chinese National Standard (CNS) A4 (210X 297 mm) 593305 A7 B7 V. Invention Description (197) ^ '(Please read the notes on the back before filling this page) Employees of the Central Standards Bureau of the Ministry of Economic Affairs The cooperative prints a 4-necked, 5-liter bottomed flask equipped with a mechanical stirrer, a pressure equalization addition funnel with a nitrogen inlet tube, a thermometer, and a condenser with a bubbler. The bubbler was connected to two 2 litre soda valves via a polyethylene tube. The first was empty and was placed to prevent reversal of the substance in the second soda valve (1 litre of concentrated subacid steel solution). -Aspirate while entering the reaction vessel. Dioxin was H79.50 g; [weight was not corrected for purity, 85% purity]) was added to 3 liters of methanol in the flask. A 25% methanol-based sodium methoxide solution (64.83 g) was placed in the funnel, added dropwise, and stirred under nitrogen for 10 minutes. After the addition of the additive, the orange mixture was heated to reflux for 20 hours. After this period, 167 ml of 4N Hci was added dropwise to the reaction mixture still refluxing by the addition funnel (warning: HCN was released at this time!). The reaction mixture became pale 'to a pale kumquat. Then a condenser was used instead of the condenser &apos; and 1.5 litres of methanol was removed by distillation, while 1.5 litres of water was added to the flask via a funnel at the speed of a steamhouse. The reaction mixture was cooled to ambient temperature and extracted twice with 2.25 liters of two gas methane. 750 ml aliquots of ice-cold saturated Na C 1 solution and 1 N N a 0 Η were used to continuously wash the mixed extracts, and then washed again with saturated Na C 1. The organic layer was dehydrated with sodium sulfate overnight, filtered, and reduced in volume to approximately 250 ml. Toluene (300 ml) was added and the residual dichloromethane was stripped under reduced pressure, during which time the product began to form a white solid on the walls of the flask. The contents of the flask were cooled overnight and the solids were removed by filtration. Rinse with 250 ml of toluene and rinse twice with 250 ml aliquots of diethyl ether and dehydrate on an empty funnel to obtain 58.8 g of a white solid. The purity by HPLC is 97.3%. When the mother liquor is concentrated, get an additional 6.76 -200- This paper size applies Chinese National Standard (CNS) A4 (210X297 mm) Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 593305 A7-* -------- --- B7_____ V. Description of the invention (198) grams of product with a purity of 77.1%. Total yield, purity adjusted to 78%. Column 2 3 Shili] 1. Step · | ^ 3Β: convert 11 α, i7 Shen-diazonyl-3 -oxoprogesterone-4-w-7-7,21-dihydrogen methyl ester, r -lactone It is 17α-hydroxy-11-methyl- (methyl, fluorenyl) oxo-3-oxopregest-4-ene-7π, 21-di double hydrogen methyl ester, and r-lactone. Equipped with a 5-liter four-necked flask as in the previous example, except that no capture system was installed before the bubbler. Add 38.7 g of hydroxy hydrazone to the flask, then add 1 425 ml of dichloromethane and stir under nitrogen. The reaction mixture was cooled to -5 ° C using a salt / ice bath. Methanesulfonium trioxide (5 1.15 g, 0.447 mol) was added quickly, followed by the dropwise addition of triethylamine (54.37 g) in 225 ml of digas methane. The adjustment requires approximately 30 minutes of addition so that the temperature of the reaction does not rise to approximately 5 ° C. After the addition, stirring was continued for 1 hour, and the reaction contents were transferred to a 12-liter separation funnel. To this was added 2100 ml of digas methane. 700 ml of ice-cold in, HC1, IN, NaOH, and saturated Na C 1 aqueous solution were used to continuously rinse the solution. The aqueous rinse was mixed and back-extracted with 3 500 ml of dichloromethane. An organic rinsing solution was mixed in a 9-liter water tank, and 500 g of neutral alumina, an activity level of 11, and 500 g of anhydrous sodium sulfate were added thereto. The contents of the jar were thoroughly mixed for 30 minutes and filtered. The filtrate was taken out and dehydrated in the air to obtain a resinous yellow bubble. It was dissolved in 350 ml of dichloromethane, stirred off and 1800 ml of ether was added dropwise. The rate of addition was adjusted so that about half of the ether was added in 30 minutes. After about 750 ml has been added, the product begins to be divided into -201-i paper size applicable to the CNS A4 specification (210 ^ 297 public attack) (Please read the precautions on the back before filling this page) 1 «

1T 593305 A7 ____ B7 V. Description of the invention (199) A crystalline solid is isolated. The remaining ether was added over 10 minutes. The solid was removed by filtration and the filter cake was rinsed with 2 liters of ether and dehydrated in a 50 t emptied oven overnight to obtain 144.61 g (88%) of an almost white solid with a melting point of 149-150C. The substances prepared in this way usually have a purity of 98-99.9% (area%) from hPLc. In one process, a substance with a melting point of 153-153.5 ° C was obtained, and its purity was determined by HPLC to be 99 5 0 / 〇. Example 2 4 Plan 1: Step 3 &lt; ^: Method 8: Preparation of 17-hydroxy_3-oxoprogesterone_4,9- (1 1)-Di-7-7,21 --- Intrinsic purpose. Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page). Set up a 1-liter four-necked flask according to the second example. Add formic acid (250 ¾ liters) and acetic anhydride (62 ml) to the flask and stir under nitrogen. Potassium formate (6.17 g) was added and the reaction mixture was heated to an internal temperature of 40 using an oil bath (repeat to 70 ° C later for better results) for 6 hours. After 6 hours, the sulfonium mesylate was added and the internal temperature was raised to 00 t. Continue to heat and stir for 2 hours, then remove the solvent on a rotary evaporator in the air. The residue was stirred with 500 ml of ice-water for 5 minutes, and then extracted twice with 500 liters of an equal portion of ethyl acetate. The organic phases were mixed and washed continuously with ice-cold 250 aliquots of saturated steel chloride solution (twice), 1 n steel hydroxide solution, and then with a saturated sodium chloride solution. The organic phase is then dehydrated with sodium sulfate, filtered, and dehydrated in the air to obtain a yellow-white foam, which makes the glass misty when it comes into contact with scraping. A 14.65 g powder was formed and analyzed as 82.1% I, 74% ^ _ and 5.7% (area% determined by HPLC). ίΜ_25 -202- This paper rule money country_material (CNS) 210 &gt; &lt; 297 mm) 593305 A7 B7 V. Description of invention (200 Printed by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 1: Step 3 C: Method B ·· Preparation 1 7 α · Hydroxy-oxopregnancy 4,9 (11) -dihydrazone-7shen, 21_dihydrogen methyl ester, factory-inner cool. Install according to the example above 5 -A four-headed flask, and 223.26 g of acetic acid and 41.37 g of sodium acetate were added and stirred under nitrogen. Using an oil bath, the mixture was heated to an internal temperature of 100 ° C. The formate salt (1 2 3 · 65 g), and continue heating for 30 minutes. At the end of the period, stop heating and add 200 ml of ice water. The temperature drops to 40 ° C, and continues to disturb for j hours, then The reaction mixture was slowly poured into 5 liters of ice water in a 5-liter flask. The product was isolated as a gum oil. This oil was dissolved in liters of ethyl acetate and gasified with ice-cold saturation. The steel solution, 1] S [sodium hydroxide and finally 1 liter of saturated sodium gas solution was used for washing. The organic phase was dehydrated with sodium sulfate and filtered. The filtrate was dehydrated in the air to obtain a foam that disintegrated into gum oil. It was triturated with ether several times and finally solidified. The solid was filtered and rinsed with more ether to obtain 79.59 g of a yellow-white solid. This included 7 0 · 4% desired A 9 '11 enester i, 1 2 · 3% △ 11,12 acetonitis, 10 · 8% 7 α, 9 π-lactone 2_ and 5 · 7% unreacted Example 2 6 Scheme 1: Step 3D ·· Synthesis of 9,1 1 α-Epoxy 17 from -Hydroxy-3_oxoprogesterone_4_ 晞 -7 ", 21_dihydrogen methyl ester, r -Lactone. A 4-necked 500 ml reactor was equipped with a mechanical stirrer, condenser / foamer, thermometer, and addition funnel with nitrogen inlet tube. The reactor was charged with 83 ml of two 8.32 grams of crude methyl ester in methyl chloride and stirred off under nitrogen. To this was added 4.02 grams of divalent potassium phosphate 'followed by 12 ml of trichloroacetonitrile. External cooling water was passed through the reactor jacket Layer flow, -203 This paper size applies Chinese National Standard (CNS) A4 specification (210X297 mm) (Please read the precautions on the back before filling this page) —Dad.-, \ Γ 593305 A7 B7 V. Description of the invention (201) (Please read the notes on the back before filling out this page} and cool the reaction mixture to 8 ° C. Add 36 to the addition funnel over a period of 10 minutes. Ml of 30% hydrogen peroxide. After the addition is complete, the reaction mixture that was light yellow in the beginning almost turned into colorless. The reaction mixture was maintained at 9 ± 1 ° C throughout the addition and continued to stir; Fall overnight (23 hours in total). Dichloromethane (150 ml) was added to the reaction mixture, and the entire contents were added to about 250 ml of ice water. It was extracted twice with a 500 ml portion of horses. Rinse the mixed digas methane extract with 400 ml of ice-cold 3% sodium sulfite solution to break down any remaining peroxide. Then rinse with 300 ml of ice-cold in sodium hydroxide, 400 l of ice-cold 1 N hydrochloric acid, and finally with 400 ml of saline. The organic phase was dehydrated with tritium sulfate, filtered, and the filter cake was washed with 80 ml of dichloromethane. The solvent was removed in the air to obtain 9.10 g of crude product as a pale yellow solid. It was recrystallized from about 25 ml of 2-butanone to give 5.5 2 g of almost white crystals. Finally, it was recrystallized from acetone (about 50 ml) to obtain 3.16 g of long needle-like crystals with a melting point of 2 4 1-2 4 3 Ό. Example 2 7 Plan 1 _ · Step 3: Option 1: From 4f S (4f α), 7, to -hexadecane-1 i 丨 from-printed by the Consumer Consumption Cooperative of the Central Standard Bureau of the Ministry of Economic Affairs-10 '/ ?, 13'fluorene-dimethyl-3,5,20, -dioxospiro [furan_2 (3 ^ 1), 17 ·-[4,7] methylfluorene [17H] cyclopenta [a ] Philippine] -5, (2, H) · Nitrile to 9,11 α-Epoxy-17 α-Equil-3-oxogesterin-4-hydrazone-7α, 21-dimethanoate, r -Lactone. A diketone (20 g) was charged into a clean and anhydrous reactor, followed by 820 ml of MeOH and 17-6 ml of a 25% NaOMe / MeOH solution. The reaction mixture was heated to reflux conditions (approximately 67 ° C) for 16 to 20 hours. Li-204- This paper size applies Chinese National Standard (CNS) A4 (210X297 mm) 593305 Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7 B7 V. Description of the invention (202) 40N 4N HCH official product cold. The solvent was removed by distillation at atmospheric pressure. 100 ml of toluene was added and the residual methanol was removed by azeotropic distillation with toluene. After concentrating, the crude glycine was dissolved in 206 * L of dichloromethane and cooled to 0 ° C. Methanesulfonyl chloride (5 liters) was added followed by slowly adding 10.8 ml of triethylamine. The product was stirred for 4 5 minutes. The solvent was removed by emptying the steamed noodles to obtain the crude formate i. In a separate anhydrous reactor, 5.93 g of formic acid bell, 240 ml of formic acid 'were added, followed by 118 ml of Sf acetate. The mixture was heated to 70 ° C for 4 hours. The formic acid mixture was added to the concentrated mesylate solution L prepared above. The mixture was heated to 95-105 ° C for 2 hours. The resulting mixture was cooled to 50 ° C and the volatile components were removed by air distillation at 50 ° C. The product was distributed between 275 ml of ethyl acetate and 275 ml of water. The layer was extracted with 137 ml of ethyl acetate, washed with 240 ml of ice-cold in sodium hydroxide solution, and then washed with 120 ml of saturated NaCII. After phase separation, the organic layer was concentrated under vacuum distillation to obtain a crude ene ester. The product was dissolved in 180 ml of dichloromethane and cooled to 0 to 15 ° C. 8.68 g of dipotassium hydrogen phosphate were added, followed by 2.9 ml of trichloroacetonitrile. Over a period of 3 minutes, 78 ml of a 30% hydrogen peroxide solution was added to the mixture. The reaction mixture is stirred at 0-15 ° C for 6-24 hours. After the reaction, the two-phase mixture was separated. The organic layer was washed with 126 ml of a 3% sulfurous acid steel solution, 126 ml of a 0.5N sodium hydroxide solution, 126 ml of 1N hydrochloric acid, and 126 ml of a 10% saline solution. Dehydrate the product with anhydrous tritium sulfate, or filter on diatomaceous earth, and remove the solvent by distillation under atmospheric pressure. -205- This paper size applies to China National Standard (CNS) A4 (210X297 mm) (please Read the notes on the back before filling in this page} 1 «· 593305 A7 B7 —-— ______________ —---- — V. Description of the invention (203) Dichloromethane. The product will form crystals from methyl ethyl ketone Twice, 7.2 grams of Epileur Reynolds were obtained. 0 〇

(Please read the precautions on the back before filling in this page. Example of printing by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 2 8 Plan 1: Step 3: Option 2: No intermediates required, i, S (4, Shen), 7 , α-Hexadecyl-1 1′-Hydroxy-10 丨 Cold, 13, /? dimethyl-3f, 5,20f-trioxospiro [furan-2 (3H), 17 '/?- [4,7] Methylpyrene [17H] cyclopenta [&amp; phenanthrene] -5, (2'H) -nitrile converted to 9,11α-epoxy-17 from -Cyclo-3 -oxoprogesterone · 4-Pyrene-7-pan, 21-dihydrobiethyl ester, r, lactone. A 4-neck 5-liter round bottom flask was equipped with a mechanical stirrer, an addition funnel with a nitrogen inlet tube, a thermometer, and a There is a condenser with a bubbler attached to a sodium hypochlorite scrubber. A diketone (83.20 g) is added to 3.0 5 liters of methanol in a flask. Into an addition funnel is charged 6785 g of 25/0 ( Weight: Weight] Sodium methoxide in methanol falls. Stir under nitrogen and add the methanolate dropwise to the flask over 15 minutes. A dark orange / yellow slurry appears. The reaction mixture is heated to reflux 20 hours, and 175 ml of 4N hydrogen acid was added dropwise while continuing to reflux (alarm When this operation will be released HCN) to acess head -206! -

593305 A7 _______B7 V. Description of the invention (204) Replace the reflux condenser and remove 丨 .6 liters of methanol by distillation. At the same time, add 16 liters of 10% chlorine dropwise through the funnel at a rate compatible with the distillation rate. Aqueous sodium solution. The reaction mixture was cooled to ambient temperature, and extracted twice with a gaseous distillate of 2.2 $ ^ aliquots. Rinse the combined extracts with ice-cold 750 ml aliquots of 1 N sodium hydroxide and saturated sodium chloride solution. The organic layer was dehydrated by azeotropic distillation with methanol at atmospheric pressure to a final volume of 1 liter (the entire 0.5% was removed for analysis). Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) Add the concentrated organic solution (based on esters) back to the installation as before, but without the original response of the H CN trap In the flask. The flask was cooled to 30.7 g of methanesulfonium, and stirred under nitrogen. The addition funnel was charged with 3,2.5 grams of triethylamine, which was added dropwise over a period of 5 minutes, keeping the temperature at 5 ° C. Stirring was continued for 2 hours while warming the reaction mixture to ambient temperature. Prepare a column containing 250 grams of Dowex 50 W X 8.100 acid ion ion exchange resin and rinse with 250 ml of water, 250 ml of methanol, and 500 ml of digas methane before use. The reaction mixture was allowed to flow down the column and collected. Prepare a new column and repeat the previous procedure. A third 250-gram column containing Dowexl X 8-200 basic ion-exchange resin was prepared and pre-treated as in the case of the acid resin described above. The reaction mixture was allowed to flow down the column and collected. A fourth test resin column was prepared, and the reaction mixture was again flowed down the column and collected. Pass each column, followed by flushing the column with 250 ml of digas methane. Each pass takes approximately 10 minutes. The solvent rinsing solution was combined with the reaction mixture, and the volume was reduced to about 500 ml under emptying, and 2% was removed for quality management. The residue was reduced to a final volume of 150 liters (crude mesylate solution). -207- This paper size applies Chinese National Standard (CNS) A4 specification (210X297 public attack) &quot; &quot; &quot; 960 ml of formic acid, 472 ml of acetic anhydride and 2 3 · 70 g of potassium formate were added to the original 5-liter reaction apparatus. Stir under nitrogen and heat the mixture to 70 ° C for 16 hours. Then increase the temperature to 100 ° C, and the crude methanesulfonate solution was added via the drain over 30 minutes. Since dichloromethane was distilled out of the reaction mixture, the temperature dropped to 8 5 ° C. After it had been completely removed After that, the temperature was ramped back to 100 ° C and maintained for 2.5 hours. The reaction mixture was cooled to 40 ° C and the formic acid was removed under pressure until the minimum stirring volume (about 150 ml) had been reached ). Cool the residue to ambient temperature 'and add 3 7 5 ml of dichloromethane. Rinse the diluted residue with ice-cold 1 liter saturated sodium emulsified solution, 1 N carbon steel and rinse again. Rinse with vaporized sodium solution. The organic phase was dehydrated with magnesium sulfate (150 g) and filtered to obtain a dark reddish brown solution (crude methyl ester solution). A 4-necked 1-liter reactor was equipped with a mechanical stirrer and condensed / Bubbler, thermometer and addition funnel with nitrogen inlet tube. The reactor was charged with a crude olefinic ester solution (estimated at 60 g) in 600 ml of digas methane 'and under nitrogen Break; add 24.0 grams of divalent scaly acid section, and then add 87 7 liters of trigas acetonitrile. Pass external cooling water through the jacket of the reactor and cool the remote reaction mixture to 10 ° C. In Add 147 ml of 30% osmium peroxide to the addition funnel and mix within 30 minutes. After the addition is complete, the 'initial dark reddish brown reaction mixture turns to pale yellow. During the entire addition, the reaction mixture is Maintain at 10 ° C and 1 ° C overnight and continue to stir overnight (total of 23 hours). Separate the phases and use 120 ml as a portion of the gas cylinder: extract the water-containing portion twice. Then add 2 1 〇 亳 L 3% sulfurous acid steel solution, rinse mixed organic .According to the starch / iodine test paper, there are -208- This paper size is applicable to the Chinese National Standard (CNS) M specification (21〇X 297 mm) (Please read the precautions on the back before filling this page) Order 593305 A7 —__ Β7 ____ V. Description of the invention (206) ~~ After the machine and the water-containing part do not contain peroxide, repeat this step twice. Use 2 j 〇 亳 to make 1 aliquot of 1N sodium hydroxide, 1 The organic phase was continuously flushed with N-hydrogen acid and finally washed twice with brine. The organic phase is dehydrated azeotropically to a volume of about 100 ml. Fresh solvent (250 ml) is added and distilled to a 100 ml volume in a stagnant manner. The residual The solvent gave 57.05 g of a crude product as a yellow foam in the form of a gum. A portion (51.01 g) was further dehydrated to a constant weight of 44.3 g and quantitatively analyzed by HPLC. Analyzed as 2 7.1% EPX. Example 2 9 A reaction flask was charged with 11-hydroxyandrostenedione dione (429.5 g) and toluenesulfonic acid hydrate (7.1) under nitrogen. Ethanol (2.88 liters) was charged into the reactor, and the resulting solution was cooled to 5 ° C. Triethyl orthoformate (3 3 4.5 g) was added to the solution over a period of 15 minutes at 0 to 15 ° C. After the addition of triethyl orthoformate was completed, the reaction mixture was warmed to 40 ° C and reached this temperature for 2 hours, then the temperature was raised to reflux, and the reaction was continued under reflux for an additional 3 hours . The reaction mixture was cooled under vacuum and the solvent was removed under vacuum to produce 3-ethoxyandrost-3,5-diene_ 1 7-one 0 f Example 3 0 -from 1 1 α -Lightyl Conrad Enamines (please read the notes on the back before filling this page)

Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 0 0

This paper size applies to China National Standard (CNS) A4 (210X 297 mm) Staff Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs, Yin Fan 593305 A7 — __Β7 5. Description of the invention (207) Sodium cyanide (1.72 g) In a 25-litre% neck flask equipped with a mechanical stirrer. Water (2.1 ml) was added and the mixture was stirred with heating until the solids dissolved. Add dimethylformamide (15 liters), followed by hydroxycono Reno (5.0 g). To this mixture was added a mixture of water (0.4 liters) and sulfuric acid (1.49 g). The mixture was heated to 8 52.5 hours, at which time HPLC analysis showed complete conversion to the product. The reaction mixture was cooled to room temperature. Sulfuric acid (0.83 g) was added, and the mixture was stirred for one and a half hours. The reaction mixture was added to 60 ml of water cooled in an ice bath. Rinse the flask with 3 ml DMF and 5 ml water. The slurry was stirred for 40 minutes and filtered. Rinse the filter cake twice with 40 ml of water and dehydrate in an air oven at 60 ° C overnight to produce 11-hydroxyenamine, which is 5, hydrazine (5, from), 7, 5-amino group Hexadecyl-hydroxy-10 · di, 13'α-dimethyl · 3 ', 5-dioxospiro [furan-2 (3Η), 17' from (5'11)-[7,4] sub Formamidine [411] cyclopentane | &gt;] phenanthrene] -5, -nitrile (4.9g). Example 3 Conversion of 1-11 Hydroxyl Cononol to Dicyto

-210- This paper size applies to Chinese National Standard (CNS) Α4 specification (210X297 mm) (Please read the precautions on the back before filling this page)

593305 A7 Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs B7 V. Invention Description (208) Flask. Add water (1.26 ml) and heat the flask slightly to dissolve the solid. Dimethylacetamide [or dimethylformamide] (9 liters) was added, followed by 11 π-hydroxyconreno (3.0 g). In a reaction flask, a mixture of sulfuric acid (0.47 g) and water (0.25 ml) was added, while sandalwood was added. The mixture was heated to 95 ° C for 2 hours. HPLC analysis indicated that the reaction was complete. Sulfuric acid (0.27 ml) was added and the mixture was stirred for 30 minutes. Introduce additional water (25 ml) and sulfuric acid (0.90 ml) and stir the reaction mixture for 16 hours. The mixture was then cooled to 5-10 ° C in an ice bath. The solid was separated by filtration through a frit glass filter, followed by washing twice with water (20 liters). The solid diketone is 4 丨 S (4 丨 sink), 7 丨 sink · hexadecane_u 丨 hydroxyl-10′々, 13′lu-dimethyl-3 ′, 5,2〇,- Dioxospiro [furan-2 (311), 44,7] methylpyrene [17Η] cyclopenta [a] phenanthrene] -5, Lu (2, Η) _nitrile, dehydrated in a vacuum oven to produce 3 0 g of solid. Example 3 2 A suspension of 5.0 g of diketone produced in the method described in Example 31 in methanol (100 ml) was heated to reflux, and 25% methanol (5.8 Halo) NaOH's sodium methoxide solution. The mixture became homogeneous. After 15 minutes, a precipitate appeared. The mixture was heated to reflux and became homogeneous again after about 4 hours. Continue heating to reflux for a total of 23.5 hours and add 4.0 n HC1 (1 () ml). A total of 60 ml of hydrogen cyanide solution in methanol was removed by distillation. Water (57 ml) was added to the distillation residue f within 15 minutes. During the addition of water, the pH of the solution rose to 81.5 C, and an additional 4 ml of hydrogen cyanide / methanol solution was removed by distillation. After the addition of water is complete, the admixture becomes a cloud 211-

(Read the precautions on the back before filling in this page) 1 «· Order p i n am A7

593305 5. Description of the invention (209) and remove the heat source. The mixture was stirred for 3.5 hours while the product slowly formed crystals. The suspension was filtered, and the collected solid was washed with water, dehydrated on the funnel with i gas vapor, and dehydrated at 9 2 ° C (2 mer mercury) for 1 6 J, 2 · 9 8 Gram _ white solid. The solid was 9 1.4% of a light-based ester, that is, 11 α, 17α_dihydroxy_3_oxopregnene-7 precipitate, 2 methyl hydrogen dicarboxylate, and lactone. The yield was 56.1 〇 / 〇 . Example 33 The diketone prepared in the manner described in Example 31 was charged into a clean, anhydrous 3-necked reaction flask equipped with a thermometer, a Dean-Stark trap, and a mechanical stirrer. Methanol (24 ml) was charged into the reactor at room temperature (2 2. (:) and the resulting slurry was stirred for 5 minutes. 25 weight ❻ / 〇 in methanol (5 2.8 pen liters) Sodium methoxide solution was charged into the reactor, and the mixture was stirred at room temperature for 10 minutes, during which time the reaction mixture became a light brown clear solution, and a slight exotherm (2_3 ° C) was observed. Control The rate of addition avoids the temperature of the bottle exceeding 30 ° C. The mixture is then heated to reflux conditions (approximately 6 7 C) 'and maintained at reflux for 16 hours. Then samples are taken and the conversion is analyzed by Η PLC The reaction is continued under reflux until the residual diketone does not exceed 3% of the diketone loading. During the reflux, the 4N HC1 (120 ml) filled in the reaction bottle, resulting in the generation of HCN, can be in the scrubber Stop it. After the reaction is completed, 90-95% of the methanol solvent is distilled out of the reaction mixture at atmospheric pressure. During the distillation, the temperature of the liquid surface changes from 6 7 -7 5 ° C, and at Before disposal, treat with caustic and bleach containing η c N Distillate. After removing the methanol, cool the reaction mixture to room temperature. • 212- This paper size applies Chinese National Standard (CNS) A4 specification (21〇 &gt; &lt; 297mm) (Please read the Note: Please fill in this page again) 丨 «Printed by 1T Printed by the Consumers 'Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 593305 Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economics A7 __________B7 V. Description of the invention (210) When the mixture is cooled to 40- At 45 ° C, the solid product began to precipitate. At 25 ° C, if necessary, an aqueous solution containing 5% by weight sodium bicarbonate (12,000 liters) was charged into the cooled slurry, and the obtained slurry was then charged. The mixture was cooled to 0 C for about 1 hour. The treatment of bicarbonate steel can effectively remove the unreacted diketone from the reaction mixture. Stir the slurry at 0 ° C for 2 hours to complete the precipitation and crystallization. The solid product was then removed by filtration, and the filter cake was rinsed with water (100 ml). The product was dehydrated to constant weight at 80-90 ° C at 26 ° C, with mercury emptied. After dehydration, the water Content is less than 0 2 5 weight 0 / 〇. The adjusted Mohr yield is about 7 7-80 weight%. Example 3 4 The diketone (1 equivalent) and sodium methoxide (4 · 8 equivalent) prepared according to Example 31 were prepared in methanol. The solvent is reacted in the presence of zinc iodide (1 equivalent). According to the extraction process described herein, or where digas methane extraction is excluded, brine and caustic washing, and non-extraction of the sodium sulfate dehydration step Process to treat the reaction product. In the non-extraction process, a 5% by weight sodium bicarbonate solution was used in place of toluene. Example 3 5 The hydroxy ester (1 · 97 g) prepared by Example 34 was mixed with tetrahydrofuran (20 ml), and the resulting mixture was cooled to 70 ° F. Sulfachloro (0.8 ml) was added and the mixture was stirred for 30 minutes, followed by acetone (1.3 g). The reaction mixture was warmed to room temperature and stirred for an additional 2 hours. The mixture was then degassed with dichloromethane and extracted with water. The organic layer was concentrated to give crude ethyl ester (.97 g). A small sample of the crude product was analyzed by HPLCZ. The analysis shows that the ratio of 9,1 1 -alkene: 1 1,1 2 -chain 晞: 7,9 -lactone is -213- The paper size is suitable for wealth_ 家 转 (CNS) M size (2 Shu 297 public attack ) (Please read the notes on the back before filling this page) Order _ Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 593305 A7 __ _____B7___ V. Description of the invention (211) 75 · 5: 7 · 2: 17 · 3. This process is mixed into one step, introducing a release group and removing it from the 9,1 1 -chain dilute structure introduced into the fluorenyl ester, that is, the reaction is to continue the fluorenyl chloride to cause the 1 1 from the transester of the formula V from- The radical is replaced by a halide, followed by a dehydrohalogenation reaction of the structure 疋 Δ-9,11. The use of strong acids (such as formic acid) or desiccants such as acetic anhydride is therefore not used, and the formation of ene esters is completed. It also precludes another process from generating a reflux step of carbon monoxide. Example 36 6 The hydroxyester (20 g) and dichloromethane (400 ml) prepared in Example 34 were added to a clean, anhydrous three-necked round bottom flask equipped with a mechanical stirrer, addition funnel, and thermocouple. The resulting mixture was stirred at ambient temperature until a complete solution was obtained. The solution was cooled to 5 using an ice bath. (:. Methane dichloromethane (5 liters) was quickly added to the ch2C12 solution containing the hydroxy ester, and then triethylamine (10-8 ml) was slowly added dropwise. The addition speed was adjusted so that the reaction temperature The temperature was above 5 ° C. The reaction was polar exothermic; therefore cooling was required. The reaction mixture was stirred at about 5 ° C for 1 hour. When the reaction was complete (HPLC and TLC analysis), the mercury was emptied at about 0 ° C at 26 ° C. The mixture was concentrated until it became a viscous slurry. The resulting slurry was diluted with CH2C12 (160 ml), and the mixture was concentrated under approximately 2 × mercury at approximately 0 ×: to obtain a concentrate. The concentrate was found Concentrate (where R3 = H, and -AA and -B-B- are -CH2_CH2-a mesylate product of formula IV, which is 11 α, 17 dihydroxy · 3 · oxopregn-4-ene- 7 Jiang, 21 · hydrogen methyl dicarboxylate 'Γ-lactone to 17α -hydroxy- lla_ (methylsulfonyl) oxo-3 -oxoprogesterone-4 · 晞 _ 7 α, 2 1 -dicarboxylic acid Hydrogen methyl ester, -lactone) has a purity of 8 2% (Η PLC area ° / 〇). No separation is required, and 214- paper is used directly in the next reaction. Scale applicable Chinese National Standard (CNS) Α4 size (210x297 mm 1 &quot; (Please read the Notes on the back to fill out this page)

593305 A7 B7 5. Description of the invention (212) The substance. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the notes on the back before filling this page). Add potassium formate (4.7g), formic acid (16ml) and acetic anhydride (8ml, 0.084mol) to the container. Clean, anhydrous reactors with mechanical stirrers, condensers, thermocouples and heating hoods. The resulting solution was heated to 70 ×: and stirred for about 4-8 hours. The addition of acetic anhydride is exothermic and generates a gas (CO), so that the rate of addition needs to be adjusted in order to control the temperature and gas generation (pressure). The reaction time for the preparation of the active elimination agent depends on the amount of water present in the reaction (formic acid and potassium formate each contain approximately 3-5% water). The elimination reaction is sensitive to the amount of water present; if &gt; 〇 · 丨 〇 / ❶ water (K F), the degree of 7,9-lactone impureness will increase. It is not easy to remove by-products from the end product. When KF &lt; 0.1% water is shown, the active elimination agent is moved to the mesylate concentrate (0.07 mol) prepared in the previous step. The resulting solution was heated to 95 C 'and the volatiles were evaporated off and collected in a Dean Stark trap. When the evolution of volatile substances ceases, replace the Dean Stark trap with a condenser and heat the reaction mixture to 9 5 additional 1 hour ° When completed (Ding 1 ^ and 11? 1 ^ (: analysis &Lt; 0.1% starting material), the contents were cooled to 50 ° (:, and empty distillation was started (26 as mercury / 50. (:). The mixture was concentrated to a viscous slurry and then cooled To ambient temperature. The resulting slurry was diluted with ethyl acetate (137 ml), and the solution was stirred for 15 minutes, then diluted with water (137 ml). The layers were separated and re-extracted with ethyl acetate (70 ml). The lower liquid layer. Rinse the mixed ethyl acetate solution once with saline solution (20 ml) and rinse twice with ice-cold 1 NN a 0 Η solution (120 ml each). Measure the pH of the liquid layer Alas, if the remaining rinse solution ρ η 値 &lt; 8, the organic layer is rinsed again. When the remaining rinse solution ρ Η 値 &gt; 8 is used, the salt solution -215- G's Zhang scale applies the Zhongguanjia standard (CNS) M specifications (GX297 public director) 593305 Α7 Β7 V. Description of the invention (213) Liquid (120 ml) Rinse the ethyl acetate layer once and use a water bath at 50 ° C to concentrate it to dryness by rotary evaporation. The resulting enester, solid product, is 17 π-hydroxy-3-oxoprogesterone-4,9 ( 11) -Diene-7 α, 21 · dihydrogen methyl ester, lactone, weighing 92 g (77 mol 0/0 yield). A hydroxy ester (1000 g; 〇 · 2 2 mol) into a 2-liter 3-necked round bottom flask equipped with a mechanical stirrer, addition funnel and thermocouple 'using a circulating cooling bath with automatic temperature control. The flask was dehydrated before the reaction Because methanesulfonyl chloride is sensitive to water. Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the precautions on the back before filling this page). Into the flask, and dissolve the hydroxyester therein with stirring. The solution was cooled to 0 ° C., and the formazan enzyme chloride (25 ml; 0.32 mol) was charged into the flask via an addition funnel. The funnel was charged with triethylamine (50 ml; 0.59 mol) into the reactor with additional dichloromethane: f e (34 ml) rinse the funnel. The addition of triethylamine is highly exothermic. The addition time under stirring and cooling is about 10 minutes. The charged mixture is cooled to 0 ° C and maintained at this temperature. During the additional 45 minutes, during this period, the liquid surface space of the reaction flask was filled with nitrogen. Then, a sample of the reaction mixture was analyzed by thin layer chromatography and high-efficiency liquid chromatography to check the reaction. Finished. The mixture was then stirred at 0 ° C for an additional 30 minutes, and the reaction was checked for 10% again. When analysis showed that the reaction was substantially complete; the solvent was removed at 0 ° C under 26 "mercury Two-gas methane stripping. Gas chromatography analysis of the distillate indicated the presence of methanesulfonyl chloride and triethylamine. Subsequently, digas methane (800 ml) was charged into the reactor, and the resulting mixture was stirred at a temperature range of 0 to ΐ5π for 5 minutes. Once again -216-

Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 593305 A7 B7 V. Description of the invention (214) The solvent was stripped under 26 &quot; mercury air, and 113 was 11, -AA- and -BB-was-CH2-CH2_ And R1 is a mesylate salt of formula IV. The purity of this product is about 90-95% by area. To prepare the elimination agent, potassium formate (23.5 g; 0.28 mol), formic acid (80 ml) and acetic anhydride (40 ml) were mixed in a separate anhydrous reactor. Formic acid and acetic anhydride were pumped into the reactor and maintained at a temperature above 40 ° C during the addition of acetic anhydride. The elimination agent mixture was heated to 70 ° F to remove water from the reaction system. This reaction was continued until the moisture content was determined to be less than 0.3% by weight based on the F i s hr analysis. The elimination solution is then transferred to a reactor containing a concentrated crude potassium sulfonate solution prepared as described above. The resulting mixture was heated to a maximum temperature of 95 ° C and the volatile distillates were collected until no more volatiles were produced. Distillation was stopped at about 90 ° C. When the distillation was complete, the reaction mixture was stirred at 95 ° C for an additional 2 hours, and the completion of the reaction was checked by thin layer chromatography. When the reaction was complete, the reactor was cooled to 50 ° C and the formic acid and solvent were removed from the reaction mixture at 50 ° F, 26 ° C, and 100% mercury. The concentrate was cooled to room temperature 'followed by introduction of ethyl acetate (688 ml), and the mixture of ethyl acetate and the concentrate was stirred for 15 minutes. A 12% saline solution (6.88 ml) was introduced at this time to assist in removing water-soluble impurities from the organic phase. The phase was then allowed to settle for 20 minutes. The liquid layer was transferred to another container and charged with an additional amount of ethyl acetate (350 ml). The back extraction of the liquid layer was completed within 30 minutes, then the phase was allowed to settle and mixed with the ethyl acetate layer. The mixed ethyl acetate layer was charged with a saturated sodium chloride solution (600 ml), and was stirred for 30 minutes. The phase was then allowed to settle. Remove the liquid layer. Completion amount -217- This paper size applies to Chinese National Standard (CNS) A4 specification (210X297 mm) (Please read the precautions on the back before filling this page)

593305 Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs A7 B7 V. Description of the invention (215) Rinsing effect of sodium carbonate (600 ml). The organic phase was separated from the second remaining rinse. The organic phase was then rinsed with 1N sodium hydroxide (600 liters) with stirring for 30 minutes. The phase was allowed to settle for 30 minutes and the liquid layer was removed. Check the p 层 of the liquid layer and find it &gt; 7. Rinse even further for 15 minutes using saturated chlorinated steel (600 ml). Finally, the organic phase was concentrated at 50 x: 2 6 ', mercury was emptied, and the product was recovered by filtration. After dehydration, the final product was a foamy brown solid. Further dehydration for 24 hours at a reduced pressure of 45 ° C yielded 95.4 g of an enester product, which was 68.8% analyzed. For the first few esters and the last enester, the modified Moire yield was 74.4%. Example 3 8 The procedure of Example 37 was repeated, except that the multiple rinse step was eliminated by treating the reaction solution with an ion exchange resin. Basic alumina or alkaline dioxide. The conditions for treatment with basic silica are listed in Tables 38 and 8. It was found that each of these treatments effectively removed impurities without requiring the multiple flushing effect of Example 44. -218- This paper size applies to Chinese National Standard (CNS) A4 (210X297 mm) (Please read the precautions on the back before filling this page)

593305 A7 B7 V. Description of the invention (216 factor oxidative oxidative factor given 2 g / 125 g of product Table 3 8) * The purpose of the test is to treat the reaction mixture with experimental alumina in order to remove the Et3N · HC1 salt and exclude IN The key result of washing with NaOH and 1NHC1 is 93% yield. 2 g / 125 g of oxidative silica. The reaction mixture was treated with cheaper alkaline silica to remove the Et3N · HC1 salt and exclude 1N NaOH and The washing output of 1NHC1 is 95% (Please read the precautions on the back before filling this page)

Example 39 In a 100-liter reactor, potassium acetate (4 g) and trifluoroacetic acid (42.5 ml) were mixed. Trifluoroacetic anhydride (9.5 ml) was added at a rate that controlled the temperature below 30 ° C during the addition. The solution was then heated to 30 C30 minutes to provide a scavenger that could be used to convert the formate salt of formula IV to the enolate of formula 11. The pre-formed TF A / TF A anhydride eradicating agent was added to the previously prepared mesylate solution of Formula 1V. The resulting mixture was heated to 40 C 4 · 5 hours' for a period of T L · C or HP L · C to check the degree of conversion. When the reaction was complete, the mixture was transferred to a 1-necked flask and concentrated under reduced pressure at room temperature (2 2 C) to dryness. To this mixture was added ethyl acetate (137 ml) to obtain complete dissolution of the solid phase material, followed by the addition of a water / brine mixture (137 ml) and the resulting two-phase mixture was stirred for 10 minutes. Then Gu Que phase points -219- This paper size applies Chinese National Standards (CNS) A4 specifications (210'〆297mm) Ordering economy, 郅 Central Bureau of Standards and Staff Consumption Cooperation 衽 Printing 593305 Staffing of Central Standards Bureau of Ministry of Economic Affairs Cooperative printed A7 B7 V. Description of invention (217) 20 minutes away. The strength of the brine was 24% by weight. The liquid phase was brought into contact with an additional amount of ethyl acetate (68 liters) and the two-phase mixture thus prepared was stirred for 0 minutes' and then allowed to stand for 5 minutes to separate the phases and the mixture was obtained from two extractions The ethyl acetate layer was 2 to 4 by weight. / () Of saline (20 ml), another aliquot of 24% by weight saline (60 ml), 1N sodium hydroxide solution (150 ml), and another portion of saline (60 ml) rinse. After each liquid phase was added, the mixture was stirred for 10 minutes and allowed to stand for 15 minutes to separate it. Under reduced pressure, the resulting solution was concentrated to dryness using a water aspirator at 45 ° C. The solid product (8.09 g) was analyzed by HP LC, and it was found to include 8 3 · 4 area% of fluorenyl ester, 2 · 4 5 area% of 1, 1, 2 -alkene, 1 · 5% of 7, 9 -Lactone and 1.1 0/0 unreacted mesylate. Example 40 0 A mesylate (1.0 g), isopropylacetate (10 g) and p-toluenesulfonic acid (5 mg) having the structure prepared in the previous Example 23 were placed in a 50 ml flask Medium and heat to 90 ° C and stir. After 5 hours, the mixture was cooled to 25 ° C and concentrated in the air under 10 mm Hg. Dissolve the residue in CH2C12 (20 mL) and hydrate with 5% water.

NaHC03 rinse. The ch2C12 layer was concentrated in the air to obtain 1.47 g of a yellow-brown oil. This material was recrystallized from CH2C12 / Et20 to obtain 0.50 g of enol acetate of formula IV (Z). This material was added to a mixture of steel acetate (0.12 g) and acetic acid (2.0 ml) previously heated to 100 ° C and stirred. After 60 minutes, the mixture was cooled to 25 ° C and diluted with dChp (0 ml). The solution was rinsed with water (20 ml) and dried over MgS04. Remove dryness by filtration -220- This paper size applies to Chinese National Standard (CNS) A4 (210X 297mm) (Please read the precautions on the back before filling this page)

Employees' Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs, Yin Fan 593305 A7 B7 V. Description of the invention (218) agent, and the filtrate was concentrated in the air to obtain 0.4 g of the desired 9,1 1 -chain, IV (Y). The crude product contained less than 2% of 7,9-lactone impurities. Example 4 Thermal elimination of 1-mesylate in D M S Ο

A mixture of 2 g of mesylate and 5 ml of D M S 0 in the flask was heated to 80 ° C for 22.4 hours. HPLC analysis of the reaction mixture showed that no starting material was detected. To the reaction was added water (10 ml), and the precipitate was extracted three times with dichloromethane. The mixed dichloromethane layer was washed with water, covered with magnesium sulfate, dehydrated, and concentrated to obtain an olefinic ester. Example 4 2 In a 50¾ liter pear-shaped flask, a stirring solid of the formula η a 晞 ester (1.07 g, analysis containing 74.4% fluorenyl ester), trichloroacetamide (0.32 g) and Dipotassium hydrogen phosphate (0.70 g) was mixed with digas methane (15.0 ml.) To obtain &amp; clear solution. Hydrogen peroxide (30% by weight; 5 · 〇) was added over a period of 1 minute via a pipette.亳 liter). The resulting mixture was stirred at room temperature for 6 hours. At this time, PLC analysis showed that in the reaction mixture, the ratio of epoxy mexrenol to ethyl ester was about 丨: 1. In this reaction mixture Add extra -221- This paper size applies to China National Standards (CNS) A4 (2mm) x 297mm 1 ~ — (Please read the precautions on the back before filling this page}-Order 593305 Central Bureau of Standards, Ministry of Economic Affairs A7 B7 printed by employee consumer cooperatives ----- ~~ ------ ^ V. Description of the invention (219) Trichloroacetamide (0.3 2 g), and continue to react for more than 8 hours under stirring, after which the remaining ethyl ester is displayed The proportion has been reduced to 10%. Add additional trichloroacetamide (0.88 g) and allow the reaction The mixture was allowed to stand overnight, at which time only 5% of the unreacted ene ester was left relative to the epoxy mexenolide in the mixture. Example 4 3 A vinegar of formula 11A (5.4 g; analysis containing 7 4.4) % Ethyl ester) was added to a 100 ml reactor. To this ene ester were added trichloroacetamide (4.9 g) and dipotassium hydrogen phosphate (3.5 g) both in solid form, followed by Dichloromethane (50 ml). The mixture was cooled to 15 ° C and 30% hydrogen peroxide (25 g) was added over a period of 10 minutes. The reaction mixture was allowed to return to 20 ° C, and Stir at this temperature for 6 hours, at which time the conversion was checked by HPLC. The remaining enester was determined to be less than 1% by weight. The reaction mixture was added to water (100 ml), and phase separation was allowed, The methane layer was removed. Sodium hydroxide (0.5 N; 50¾ liters) was added to the dichloromethane layer. After 20 minutes, phase separation was allowed, and HC1 (0.5N; 50 ml), then phase separation was allowed, and the organic phase was washed with saturated saline (50 ml). Dehydrate with anhydrous sulphuric acid and remove the solvent. A white solid (5.7 g) was obtained. The aqueous sodium hydroxide layer was acidified and extracted, and an additional 0.2 g of the product obtained by the extraction treatment. Epoxime The yield of Xrenox was 90.2%. Example 4 4 The methyl ester of formula 11A was converted to epoxy Mexray in the manner described in Example 43, but with the following differences: The initial loading included 晞Purpose (5.4g-222- This paper size applies to Chinese National Standard (CNS) A4 specifications (210X297mm) f Please read the notes on the back before filling out this page j-order-593305 Employee Consumer Cooperatives, Central Standards Bureau, Ministry of Economic Affairs A7 B7 was printed. 5. Description of the invention (220). Analysis of 74.4% of enols), trichloroacetamide (3.3 g) and dipotassium hydrogen phosphate (3.5 g). Add hydrogen peroxide solution (12.5 ml). The reaction was run overnight at 20 ° C. Subsequent HPLC showed that 90% of the fluorene esters were converted to epoxymaxex Renault. Additional trichloroacetamide (3.3 g) and 30% hydrogen peroxide (5.0 ml) were added and the reaction was carried out for an additional 6 hours. At this time, based on the loading of methyl ester, only 2% of the residual Ester. After treatment as described in Example 43, 5.71 grams of epoxy Mexanol was produced. Example 4 5 Using the general method described in Example 43, the enol of formula IIA was converted to epoxy mexrenol. In the reaction of this example, the loading of fluorenyl ester is 5.4 (analytical content of 74.4% of fluorenyl ester), the loading of trichloroacetamide is 4.9 g, the loading of hydrogen peroxide is 25 g, and dipotassium hydrogen phosphate The loading capacity is 3.5 grams. The reaction was carried out at 20 C for 18 hours. Residual ethyl ester was less than 2%. After the treatment, 5.71 g of epoxy Mex Renault was produced. Example 4 6 The fluorene ester of formula 11A was converted to epoxy mexanol in the manner described in Example 43. The reaction temperature in this example was 28 ° C. The loadings of the substances in the reactor included enester (2.7 g), trichloroacetamide (2.5 g), dipotassium hydrogen phosphate (1.7 g), hydrogen peroxide (17.0 g) and dichloromethane (50 g) Rise). After 4 hours of reaction, the unreacted fluorene ester was only 2% based on the fluorene ester loading. After the treatment as described in Example 43, 3.0 g of epoxy Mexanol was obtained. Example 4 7 Dissolve the associative ester of formula 11A (17 g; analysis contains 72% fluorenyl ester) dissolved in dichloro-223-This paper is in accordance with Chinese National Standard (CNS) A4, description f 辂, (Please read first Note on the back page, please fill in this page} 、 Yinding Ding 593305 A7 B7__ V. Description of the invention (221) Methane (150 ml), then add trichloroacetamide (1 4 · 9 g) under slow stirring ). The temperature of the mixture was adjusted to 25 ° C, and the potassium dibasic phosphate was stirred in water (10.6 ml) in a solution of the acetamyl esterase with stirring at 400 rpm. (10.6 g) solution. Hydrogen peroxide (30% by weight solution; 69.4 ml) was added to the enzymatic / phosphoric acid / trichloroacetamide solution over a period of 3-5 minutes. No exotherm or evolution of oxygen was observed. The reaction mixture thus prepared was stirred at 400 rpm for 1 8.5 hours at 25 ° C. No evolution of oxygen was observed throughout the reaction. Dilute with water (69.4 ml) The reaction mixture, and the mixture was stirred at about 250 rpm for 15 minutes. No temperature control was required for this operation, Generally operates at room temperature (any temperature in the range of 5-2 5 ° C is acceptable). The liquid layer and the organic layer are allowed to separate, and the lower two-gas-fired layer is removed. Central Ministry of Economic Affairs Standard Bureau staff consumer cooperatives Yinfan (please read the precautions on the back before filling this page) at 250 rpm stirring rate, back extract the liquid layer with methane (69.4 ml) for 15 minutes. The layers are allowed to separate, The lower dichloromethane layer was removed. The liquid layer (177 g; pH = 7) was measured for hydrogen peroxide. The results (12.2%) showed that only 0.004 was consumed in the reaction. Moore's hydrogen peroxide is 0. 0 3 0 7 Moore's alkene. Using a small volume of two-gas methane reaction extraction is sufficient to determine that no epoxy mexanol is lost in the liquid layer. The result was confirmed by the two-gas methyl alcohol extraction, and only the three-gas acetamide was recovered in the extraction. The mixed dichloromethane solution obtained from the above extraction was mixed and mixed at a rate of about 250 rpm at 3% by weight sodium sulfite solution (122 ml) rinse for at least 15 minutes At the end of the fall period, a negative starch iodine test was observed (KI test paper; no color was observed; in a positive test, the purple table-224-I paper size was applicable to China National Standard (CNS) A4 specification (210X297 (Mm) ~ '' 593305 A7 B7 V. Description of the invention (222) shows the existence of peroxide). Allow the liquid layer and the organic layer to separate and remove the lower methylene chloride layer. Discard the liquid layer (pH = 6) . Note that the addition of sodium sulfite solution can cause a slight exotherm so that it should be added under temperature control. At a rate of about 250 rpm and at a temperature range of 15-25 t (pH = 12-13), the digas phase was washed with 0.5N sodium hydroxide (61 ml) for 4 5 minutes. Amine derived from trichloro-acetyl remove impurities in the process. The alkaline aqueous fraction was acidified, followed by extraction with dichloromethane, and it was confirmed that a small amount of% oxmex Reynolds was lost during the operation. At a stirring rate of 250 rpm, in the temperature range of i5-25 ° C, rinse the digas methane phase with 0.1N hydrochloric acid (61 liters)! 5 minutes. The layers were then allowed to separate, the lower methylene chloride layer was removed, and rinsed again at 1050% by weight aqueous sodium chloride (61 ml) at a temperature range of 25 rpm and 15-25 C. minute. The layers are again allowed to separate and the organic layer is removed. The organic layer was filtered through a pad of Solkafloc and evaporated to dryness under reduced pressure. Use a water bath temperature of 65 ° C to completely dehydrate it. An off-white solid (17.9 5 g) was obtained and analyzed by HPLC. The analysis contained epoxy Mexyl Reynolds 66.05%. The adjusted Mohr output for this reaction is 9 3 · 10/00 printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page). Dissolve the product in hot methyl Ethyl ketone (189 ml) and the resulting solution was distilled at atmospheric pressure until 95 ml of the ketone solvent had been removed. The product crystallized when the temperature was reduced to 50 ° C. Stir off at 50 ° C for 1 hour. Then reduce the temperature to 20-25 ° C and continue stirring for another 2 hours. Filter the solids and rinse them with MEK (24 ml), and dehydrate the solids to a constant weight of 9.98 g -225- This paper size applies to China National Standard (CNS) A4 specifications (210 × 297 mm) 593305 Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economy Preparation of A7 B7 V. Description of the invention (223). By HPLC analysis, it contained 93.63% epoxy Mexanol. The product was redissolved in hot MEK (106 ml) and the hot solution was filtered under pressure through a 10 micron linear filter. Rinse with an additional 18 ml of MEK and distill the filtered MEK solution at atmospheric pressure. Until 5 3 ml of solvent has been removed. The temperature was reduced to 50 ° C, at which time the product formed crystals; and it was continuously stirred at 50 t for 1 hour. The temperature was then lowered to 20-2 5 C 'and stirring was continued for another 2 hours at this temperature. The solid product was filtered and rinsed with MEK (18 mL). The solid product was dehydrated to a weight of 8.32 g, and was analyzed by quantitative HPLC, and contained 99.6% epoxy mexrenol. Finally, less than 1.0% was lost during dehydration. According to the reaction and treatment in this example, the total production of epoxy Mexileau is 65. 8%. This total yield reflects a reaction yield of 93%, an initial crystallization recovery of 78.9%, and a recrystallization recovery of 89.5%. Example 4 8-Formula IIA utilizes cyclic argonization of toluene In the general method described in Example 46, the alkenyl ester of formula na was converted to apirenaline, except that toluene was used as a solvent. Loading capacity of the substance into the reactor, including methyl esters (2.7 grams), trigas acetamide (2.5 grams), dipotassium hydrogen phosphate (1.7 grams), hydrogen peroxide (170 grams) And toluene (5 () ml). The reaction was allowed to exotherm to 28 ° C and was completed in 4 hours. The resulting three-phase mixture was cooled to 15 ° C, filtered, rinsed with water, and dehydrated in the air to obtain 2.5 g of product. Example 4 9-9 · 1 1 -dienone% i oxidation in a one liter 3-necked flask will be named χνιΙΑ (where _a_a_ and -B_ B- are both -CH2-CH2- compounds χνπ " 々Compound (4067g) Soluble-226-IKEA Standard (CNS) for this paper (Please read the precautions on the back before filling this page)

593305 A7 B7 V. Description of the invention (224) Dissolved in dichloromethane (250 ml) and cooled by an external ice-salt mixture. Dipotassium phosphate (22.5 g) and trigas acetonitrile (83.5 g) were added, and the mixture was cooled to 2 ° C, and then 30% hydrogen peroxide (200 g) was slowly added over a period of 1 hour. ). The reaction mixture was stirred at 12 ° C for 8 hours' and stirred at room temperature for 14 hours. A drop of the organic layer was removed and checked for any starting fluorenone, and was found to be &lt; 0.5%. Add water (400 ml), stir for 15 minutes, and separate the layers. The organic layer was washed successively with 200 ml of potassium iodide (100%), 200 ml of sodium thiosulfate (10%), and 100 ml of a saturated sodium bicarbonate solution, and the layers were separated each time. The organic layer was dried over anhydrous magnesium sulfate and concentrated to give a crude epoxide (41 g). The product was recrystallized from ethyl acetate: digas methane to give 14.9 g of pure material. Example 5 0-Formula X V11A Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs using the epoxidation of m-peroxy benzoic acid In 250 ml of dichloromethane and cooled to 10 ° C. Under stirring, solid m-chloroperbenzoic acid (50-60% purity, 21 · 86 g) was added over a period of 15 minutes. No increase in temperature was observed. The reaction mixture was stirred for 3 hours and checked for the presence of dioxin. The reaction mixture was continuously treated with sodium sulfite (10%), sodium hydroxide solution (0.5 N), and hydrogen acid solution (5%), and finally treated with 50 ml of a saturated saline solution. After dehydration with magnesium sulfate and evaporation, 17 64 g of epoxide was obtained, which was used directly in the next step. The product was found to contain a Bayer-Villiger oxidation product, which could be removed by trituration with ethyl acetate and subsequent formation of crystals from dichloromethane. For a 500 g scale, the precipitated m-chloroperbenzoic acid was filtered according to the usual treatment. -227- This paper size applies Chinese National Standard (CNS) A4 (210X 297mm) Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 593305 A7 ____B7 V. Description of the Invention (225) Example 5 1-Composition XVIIA The ring of acetylamine was used to dissolve compound XVII A (2 g) in 25 ml of digas formamidine. Add trigas acetamide (2 g) and dipotassium phosphate (2 g). Stir at room temperature, add 30% hydrogen peroxide (10 ml), and continue stirring for 18 hours to produce peroxides (1.63 g). No Bayer Vegley oxidation products were formed. Example 5 2 Potassium hydroxide (5 6.39 g; 1 005.03 millimoles; 3,000 equivalents) was charged into a 2000 ml flask and formed with dimethyl sulfoxide (750,000 ml) at ambient temperature. Slurry. Tristrione (where R3 is Η, and _Α · Α-and-Β- Β- are -CH2-CH2-, respectively) (100.00 g; 335.01 mol; 1.0 eq.) And THF (95 6.0 ml) was charged into the flask together. Trimethylphosphonium methyl sulfate (126.14 g; 670.02 mmol; 2,000 equivalents) was charged into a flask, and the resulting mixture was heated to reflux at 80 to 85 ° C for one hour. The conversion of 17-spirooxymethylene was checked by HPLC. Approximately 1 liter of THF was stripped from the reaction mixture under vacuum, and then water (460 ml) was charged over a period of 30 minutes while the reaction mixture was cooled to 5 ° C. The resulting mixture was filtered, and the solid ethylene oxide product was washed twice with 200 ml aliquots of water. It was observed that the product was highly crystalline and could easily complete the filtration. The product was then dehydrated at 40 ° C under vacuum. 104.6 g of 3-methylenolene delta-5,6,9,11, _17_ ethylene oxide steroid product was isolated. Example 53 Under a nitrogen blanket, sodium ethoxide (41.94 g; 616.25 mmoles; 1.90 equivalents) was charged into an anhydrous 500 ml reactor. Load ethanol into the reactor-228- This paper size applies to Chinese National Standard (CNS) A4 (21 × 297 mm) (Please read the precautions on the back before filling this page)-Order 593305 Central Bureau of Standards, Ministry of Economic Affairs Employee Consumer Cooperative printed A7 V. Invention Description (226) (270.9 ml) and made copper methoxide into a slurry in ethanol. 03.90 g of malonyl malonate U; 64 8.6 8 mmoles; 2.000 equivalents) was charged into the poly I 'followed by the addition of ethylene oxide steroid U 04.60 g prepared by the method described in Example 52; 324.34 Millimoles; i.oo equivalent), and the resulting mixture was heated to reflux, that is 80-85. (: Continue heating for 4 hours, then check the completion of the reaction by H PLC. During the 30 minute period, fill the reaction mixture with water (3 3 7.86 ml) while cooling the mixture: to ^ ° C The stirring was continued for 30 minutes, and then the reaction slurry was filtered to produce a filter cake containing fine-grained amorphous powder. The filter cake was rinsed with water (200 liters each) for 2 ^, and then dehydrated under ambient air. 33 · 8 g of 3-methylenol ether △ 5,6,9,11-17-spirolactone-2 1-methoxycarbonyl intermediate was isolated. Example 5 4 3 · produced according to Example 53 Methenol ether-a ^ 7 · spironyl- 21-methoxyl intermediate (Formula Xνιπ, where R3 is η, and -A- Α-and-Β_Β-are-CH2_CH2_; 133.80 g; 313.68 millimoles; 1.0 eq.), With sodium vaporization (27.50 g; 470.52 mol; 1.50 μm), monomethylformamide (7.09 liters), and water (5 ml ) Put together into a 2000-ml reactor with caustic agitation. The resulting mixture was heated to reflux at 138-142 ° C for 3 hours, and then the reaction mixture of the reaction mixture was checked by HPlc for redundancy. Then in a period of 30 minutes, water was added to the mixture 'while the mixture was cooled to 5 X :. Stirring was continued for 30 minutes, and then the reaction slurry was filtered to recover amorphous cake-like amorphous The reaction product in a solid state. The filter cake was rinsed twice (200 liter aliquots of water), and then dehydrated. The product 3-methylenol ether_ 17 -spirolactone was dehydrated to obtain 91.6 g ( 8 2 · 3 ° / 〇Production-229- This paper standard It Finance Standard (CNS) A4 Specification (2 ^ 1 ^ Dong) (Read the precautions on the back before filling in this page)

593305 Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs A7 B7 V. Amount of Invention Description (227); 96% Analysis by Area). Example 5 5 Methanol ether (9 丨 · 60 g; 2 58 · 36 mmol; 1.0 equivalent) produced according to Example 5 4, ethanol (250 ml), acetic acid (250 ml) Water and water (250 liters) were charged into a 2000 ml reactor, and the resulting slurry was heated to reflux for 2 hours. Over a period of 30 minutes, water (600 ml) was charged while the reaction mixture was cooled to 15 ° C. The reaction slurry was then filtered and the filter cake was washed twice with water (200 ml aliquots). The filter cake was then dehydrated; 84.4 grams of the product 3-ketone △ 5,6,9,1 1-1 7-spirolactone (a compound of formula XVII, where R3 is fluorene and -A_A _ * _ b_b_ is _Ch2_CH2-; yield 9 5.9%) Example 5 6 Compound XVIIA (1 kg; 2.81 moles) was charged together with carbon tetrachloride (3_2 liters) into a 22 liter 4-neck flask. N-bromo-succinimide (538 grams) was added to the mixture, followed by acetonitrile (3.2 liters). The resulting mixture was heated to reflux and maintained at a reflux temperature of 68 ° C for about 3 hours, resulting in a clear orange solution. After heating for 5 hours, the solution turned dark. The heat source was removed after 6 hours and the reaction mixture was sampled. The solvent was stripped under vacuum and ethyl acetate (6 liters) was added to the residue remaining on the bottom. The resulting mixture was stirred, then a 5% sodium bicarbonate solution (4 liters) was added, and the mixture was stirred for 15 minutes, after which time the phase was allowed to settle. The liquid layer was removed and a saturated saline solution (4 liters) was introduced into the mixture, followed by stirring for 15 minutes. The phases were separated again, and the organic layer was stripped under the air, resulting in a thick slurry. Then add dimethylformamide (4 liters), and continue stripping to a tank temperature of 55 -230- This paper size applies to China National Standard (CNS) A4 specifications (210'〆297 mm) (Please read the back first (Notes on this page, please fill out this page) Ding 593305 A7 B7 V. Description of the invention (228) C. Allow to stand still at the bottom overnight and add daBCO (330 g) and lithium bromide (243 g). The mixture was then heated to 70X :. After one and a half hours of heating, remove the liquid chromatography sample and heat it after 3.5 hours, add additional DABCO (40 g). After 4.5 hours heating ′ water (4 liters) was introduced and the resulting mixture was cooled to 15 ° C. The slurry was filtered and the filter cake was rinsed with water (3 liters) and dehydrated on the filter overnight. The wet cake (978 g) was returned to a 22-liter flask and dimethylformamide (7 liters) was added. The mixture thus produced was heated to 105 ° C, at which point the filter cake was completely dissolved in the solution. Remove the heat source and stir off the mixture in the flask before cooling. Ice water was applied to the jacket of the reactor, and the mixture in the reactor was cooled to 14 ° C for 2 hours. The resulting slurry was filtered and rinsed twice with 2.5 l aliquots of water. The cake was dehydrated overnight under emptiness. 5 10 g of light brown solid product are obtained. Example 5 7 A 2 liter 4-necked flask was charged with 9,11-epoxy Conreno (100.00 g; 282.1 millimoles; ΐ0.00 equivalent) manufactured according to Example 4, 9, 50, or 51. ), Dimethylamidamine (650.0 ml), lithium chloride (30.00 g ·, 707.7 mmol; 2.51 equivalent) and acetone cyanohydrin (72.04 g; 77.3 ml; 846.4 mmol); 3.0 0 equivalent ). The resulting suspension was mechanically shaken with tetramethylguanidine (4 5.49 g; 49.6 ml; 395.0 millimoles, 1.40 equivalents). Then use a water-cooled condenser and a dry ice condenser (fill the dry ice in C_) to filter the system to avoid the escape of HCN. The exhaust line passes through the dry ice condenser. It enters the washing with a lot of chlorine bleach The mixture is heated to 80 ° C. -231- This paper size applies Chinese National Standard (CNS) A4 specification (210X297 mm) (Please read the precautions on the back before filling this page), τ Central of the Ministry of Economy Printed by the Consumer Bureau of the Standards Bureau Printed by the Central Consumers Bureau of the Ministry of Economy 593305 A7 B7 5. Description of the invention (229) '' After 18 hours, obtain a dark red-grass solution, cool it to the chamber, w and Stir. During cooling, sparg nitrogen into the solution to remove residual HCN using the 'exhaust line' of the bleach in the scrubber. After 2 hours, acetic acid (72 g ) Treat the solution and stir for 30 minutes. Then pour the crude mixture into ice water (2 liters) and mix it. Then treat with 10% water containing HC1 (400 liters). Overhang The mixture was stirred for 1 hour. The mixture was then filtered to obtain a dark red solid (73 g). The filtrate was placed in a 4 liter separatory funnel and extracted with methane (3x800 ml); the organic layers were mixed , And back extraction with water (2 \ 2 liters). In it it 丨 stretched and simmered in one gas; the grains' passed to 6 1 g of dark red oil. After allowing the water-containing rinses to stand overnight, it appeared Quite a lot of slaughter. The precipitate was collected by filtration and determined to be the pure product amidine (14.8 g). After dehydrating the original red solid (73 g), it was analyzed by HPLC and determined. The main ingredient is 9,11-epoxidamine. HPLC further showed that the enamine was the main component of the red oil obtained from the dichloromethane treatment. The molar yield of the amidine was calculated to be 46%. Example 5 8 According to the example 9,11 1-epoxidamine (4.60 g; 0.011261 mole; 1.00 equivalent) prepared in 5 7 was introduced into a 1000 ml round bottom flask. To this mixture was added methanol ( 300 ml) and 0.5% by weight of aqueous HC1 (192 ml), which was subsequently refluxed. 7 The subsequent removal of methanol under empty space reduced the amount of material remaining in the tank to 50 ml and caused the formation of a white precipitate. Water (100 ml) was added to the slurry, which was subsequently filtered ， Product-232- This paper size is applicable to the country of China ^^ ΐΤΤΤ〇〇297297 mm) (Please read the precautions on the back before filling out this page)-^ · 593305 A7 B7 V. Description of the invention (230) ~ Raw white Rinse the solid cake three times with water. This gave 3.747 g of the 9,1 1-epoxydione product (8 m · 3%) as a solid. Example 5 9 An epoxy diketone (200 mg; 0.49 mmol) prepared according to Example 5 8 was suspended in methanol (3 ml), and 1,8-diazine was added to the mixture. Bicyclic [5 · 4 · 〇] undecyl_7-# (DBU). When heated under reflux for 24 hours, the mixture becomes homogeneous. It was then concentrated to dryness on a rotary evaporator at 30 ° C and the residue was distributed between dichloromethane and 3.0 n HC1. The organic phase was shrunk to give a yellow solid (93 mg), which was identified to be 22% by weight of epoxy Mexanol. The yield is 20%. (Please read the notes on the back before filling this page)

Printed by the Consumers Cooperative of the Central Bureau of Standards of the Ministry of Economics of Epilele, printed by Example 60 0. To 100 mg of dione suspended in 1.5 ml of methanol, add 10 μl (0.18 equivalent) of 25% (w / w) in A solution of sodium methoxide in methanol. The solution was heated to reflux. After 30 minutes, no diketone remained and 5_ -233 appeared-This paper size is in accordance with China National Standard (CNS) A4 (210X29? Mm)

Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs 593305 A7 B7___ 5. Description of the Invention (231) Cyanoester. To this mixture was added 46 microliters of a 25% (w / w) solution of sodium methoxide in methanol. The mixture was heated to reflux for 23 hours, at which point the main product was Epileleno, as judged by ΗPLC. To 2 g of diketone suspended in 30 ml of anhydrous methanol, 0.34 ml of triethylamine was added. The suspension was heated to reflux for 4-5 hours. The mixture was stirred at 25 ° C for 16 hours. The resulting suspension was filtered to obtain 1.3 g of 5-cyanoester as a white solid. To 6.6 g of diketone suspended in 80 ml of methanol, 2.8 ml of triethylamine was added. The mixture was heated to reflux for 4 hours and stirred for 8 8 hours at 25 x during which time the product formed crystals from the solution. Filtration followed by rinsing with methanol gave 5.8 g of cyanoester as a white powder. This material was recrystallized from chloroform / methanol to obtain 31 g of a crystalline material, which was found to be homogeneous from HpLc. &lt; As can be seen from the above, it will be seen that several objects of the present invention have been achieved and other advantageous results obtained. 〃 The above composition and process can be changed in various ways without violating the scope of the present invention ', and it is intended to include all of the above descriptions and display them ^ (Please read the precautions on the back before filling this page)

-234-

593305 A7 B7 V. The events in the pictures recorded in the description of the invention (232) are to be interpreted as illustrations, without limitation. (Please read the notes on the back before filling this page) Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs -235-This paper size applies to China National Standard (CNS) A4 (210X297 mm)

Claims (1)

593305 Patent No. 08611δ698 Requested g Chinese Patent Application Replacement (September 1992) 1. A method for preparing a 3 -keto-7α_alkoxycarbonyl-substituted Δ_4,5_ steroid ', which comprises making the alkylating reagent and 4,5_dihydro. 5,7_ intrinsic steroid subject to enzymes, In the presence of a base, the lactone is substituted by a ketone or dialkoxy at the 3-carbon position of the enzyme, and further includes the following: (7). Where C (5) represents the enzyme The 5-carbon position of the steroid structure, and C (7) represents the 7-carbon position. 2. The method according to item 丨 of the patent application, wherein the type of transzyme includes 2 and or unsaturated core structures, and further includes: 9, &quot; _epoxide or its precursor; and 17 The substituent at the carbon is selected from the group consisting of spirolactone, ketone, or other spirolactone precursors. 3. The method according to item 2 of the patent claim, wherein the steroid is substituted by enzymatic and steroid-prepared steroids at the 7-7 position with a spironolactone substituent in accordance with formula XXXIII, XXXIII 〇 4. 5. 6. According to the scope of patent application! The method of the item, which includes a 17_ 鲖 structure. The method according to the scope of the patent application, wherein the lactone is enzymatic substance including a 3-alkoxy substituent. According to the method of the scope of the patent application, the method of item 5,7 593305 A8 B8 C8 D8 The base halide reacts in the presence of the test. • A method for lactone steroid compounds, which replaces the lactone, and includes the following structure: 7. — A method for preparing 4,5-dihydro-5,7_lactone steroid steroids is replaced by ketones at 3 carbons , Where C (5) represents the 5-carbon position of the steroid structure of the lactone compound, and C (7) represents the 7-carbon position, the method includes:-converting a cyano-substituted steroid to a 7-carboxylic acid, followed by The 7-carboxylic acid is reconverted to 5,7-lactone. 8. The method according to item 7 of the scope of patent application, wherein the enzymatic substance includes 3 _ iso-Δ-4,5-7-carboxy steroid, and forms including 3-dialkoxy-5,7-lactone The ketal intermediate is hydrolyzed to form 3-keto-5,7 · lactone under acidic conditions. 9. · A method for preparing a compound according to formula Π: Of which, this paper size applies to China National Standard (CNS) Α4 size (210 X 297 mm) 593305 A8 B8 C8 -A_A- represents a group _chr4_chr5 or cr4 = cr5_ R represents a lower alkoxycarbonyl or hydroxycarbonyl group of alpha (alPha); R, R, and R5 are selected from the group consisting of hydrogen, halogen, hydroxyl, low-carbon alkyl, Low carbon number alkoxy, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, cyano, aryloxy, ^-: 6- representative group &lt; 111 ^ 〇: 1111, or 1 or / 3-position Groups: R6 i ~ CH, III where R6 and R7 are selected from the group consisting of hydrogen, halogen, low-carbon alkoxy, acid group, hexaalkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxycarbonyl, Alkoxyalkyl, cyano, and aryloxy, and R and R are respectively selected from the group consisting of fluorene, fluorene, low-carbon alkoxy, fluorenyl, amido, alkoxy, fluorenyl, alkynyl, Tiger oxenyl, oxygen test, cyano, aryloxy, or R8 and R9 together contain a mosquito or heterocyclic structure, or R8 or R9 and R6 or R7 contain a carbocyclic or Heterocyclic structure, fused into a five-ring D ring, the method includes: by hydrolysis, followed by reaction with an alkylating agent in the presence of a base, alkylating a compound of formula EI, the compound of formula EI has the following structure ： -3 593305 A8 B8 C8 ______D8 6. Scope of patent application ΞΙ wherein -Α-Α-, R3, R8, R9, and -Β-Β_ are as defined above, and r 17 is (^ to (: 4 alkyl). 10. a method: IIC where -Α-Α- represents a group-CHR4-CHR5-or _cR4 = CR5 · R1 represents an alpha (lower) alkoxycarbonyl or hydroxycarbonyl group; R3, R4 and R5 are selected from the group consisting of hydrogen, Element, hydroxy, low-carbon alkyl, low-carbon alkoxy, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, cyano, aryloxy, R26 is Ci to C4 alkyl; -BB- represents a group -CHR6-CHR7_ or α _ or cardiac group: -4 593305 Where R6 and R7 are selected from the group consisting of hydrogen, halogen, low-carbon alkoxyfluorenyl, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxycarboxy, fluorenylalkyl, cyano, and aryloxy The method includes: by hydrolysis' followed by reaction with an alkylating agent in the presence of a base, the compound of formula E is alkylated, and the compound of formula E has the following structure: Wherein _A-A-, R3 and _Β_Β are as defined above, and R17 is a ^ to C 4 fe group. 11 · A method for preparing a compound according to formula E: 〇 This paper size applies to China National Standard (CNS) A4 specification (210 X 297 mm) ^ J 593305 A8 B8 C8 D8 ^^ CR5- R3, R4 and R5 are selected from the group consisting of hydrogen, halogen, hydroxyl, low-carbon alkyl, low-carbon alkoxy, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, cyano, and aryloxy, R17 is &lt; ^ to (: 4 alkyl; -BB- represents a group -CHR6-CHR7- or α- or non-azimuth group: Wherein R6 and R7 are respectively selected from the group consisting of hydrogen, halogen, low-carbon alkoxy, fluorenyl, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, alkynyl, alkoxycarbonyl, fluorenylalkyl, cyano, Aryloxy, the method includes: thermally cracking a compound conforming to formula DE 2 in the presence of an alkali metal liide, the compound of formula DE 2 having the following structure: Wherein R is Ci to C4 alkyl, and-, r17 and -B-B are as defined above. This paper size applies to China National Standard (CNS) A4 (210 X 297 mm) 593305 A8 B8 C8 D8 A method for preparing compounds according to formula DE2: Where -A_A_ represents the group _CHR, chr5_ or cr4 = cr5_ RR and R is selected from the group consisting of hydrogen, autogen, hydroxyl, low-carbon alkyl, low-mosquito alkoxy, per-alkyl, alkoxyalkyl, carbonyl , Cyano, aryloxy, R17 and R12 are respectively selected from c ^ c4 alkyl; Representative group _CHr6_chr7_ or ^ or ^ • Orientation group: Wherein R6 and R7 are respectively selected from the group consisting of hydrogen, functional elements, low-carbon alkoxy, fluorenyl, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxycarbonyl, fluorenylalkyl, cyano, Aryloxy, the method comprises: condensing a compound of formula DE 1 with a dialkyl malonate in the presence of a test, the compound of formula DE 1 having the following structure: The Chinese paper standard (CNS) applies to this paper A4 size (210 X 297 mm) 593305 8 8 8 8 A BCD 々, patent application scope 0 DE1 where -A-A-, R3, R17 and -B-B- are as defined above. 13. A method for preparing a compound according to formula DE 1: Where -AA- represents a group-CHR4-CHR5- or -CR4 = CR5- R3, R4 and R5 are selected from the group consisting of hydrogen, halogen, hydroxyl, low-carbon alkyl, low-carbon alkoxy, hydroxyalkyl, alkane Oxyalkyl, hydroxycarbonyl, cyano, aryloxy, R17 is Ci to C4 alkyl; -BB- represents the group -CHR6-CHR7- or α- or / 3-azimuth group, -8- this paper Standards apply to China National Standard (CNS) A4 specifications (210 X 297 mm) 593305 A8 B8 C8 D8 R6 and R7 are selected from the group consisting of hydrogen, halogen, low-carbon alkoxy, alkynyl, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxycarbonyl, alkoxyalkyl, and cyano Group, aryloxy group, the method includes: the formula = two times = salt recording reaction in the compound domain D wherein, R3 and the same as above 14. A method for preparing a compound according to formula d: D -9- 593305 Where -A-Λ- represents a group-CHR4-CHR5- or -CR4 = CR5- R, R and R5 are selected from the group consisting of hydrogen, halogen, light group, low carbon number alkyl group, low carbon number alkoxy group, and hydroxyalkane Group, alkoxyalkyl group, hydroxycarbonyl group, cyano group, aryloxy group, R17 is Ci to c4 alkyl group; -BB- represents group -chr6-CHR7_ or α- or / 5-azimuth group:% _ / R7 .I 1 1 CHC Η 2 CH —- III where R and R are respectively selected from the group consisting of hydrogen, halogen, low-carbon alkoxy, fluorenyl, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, and alkyl , Alkoxycarbonyl, alkoxyalkyl, cyano, aryloxy, and the reading method includes: hydrolyzing a compound of formula C to a 7 α-carboxylic acid, and reacting with trialkyl orthoformate under acidic conditions, the formula The compound of C has the following structure: R3 c wherein-Α-Α-, R3 'R17 and -Β-Β- are as defined above. -10- This paper size applies to Chinese National Standard (CNS) A4 specification (210 X 297 mm) A8 B8 C8 D8 Application scope of patent 15 · —A method for preparing compounds conforming to formula I: R3 R80 Where _Α · Α- represents the group -CHRLCHr5 or cr4 == cr5_ R3, R4 and R5 are selected from the group consisting of hydrogen, halo, hydroxyl, low-carbon alkyl, low-carbon alkoxy, hydroxyalkyl, alkoxy Alkyl, hydroxycarbonyl, cyano, aryloxy, R26 is c ^ c4 alkyl; -BB- represents the group _CHr6_chr7 • or a group of orientation: —_ C Η — ^ C Η 2 ~~ C Η — III wherein R and R7 are selected from the group consisting of hydrogen, halogen, low-carbon alkoxy, fluorenyl, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxy, Sat oxonyl, Gas group, aryloxy group, and R80 and R90 are selected from r> r9 'or r80 and r90 together to form a ketone. R8 and R9 are respectively selected from the group consisting of hydrogen, hydrogen, low carbon number alkoxy group, alcohol group, hydroxyalkane Group, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxycarbonyl, oxyalkyl, cyano, aryloxy, or ... Contains -11-593305 8 8 8 8 ABCD with rule 9 Structures with a range of 1% or a heterocyclic ring, or R8 * R9 and r6 * r7 together containing a carbocyclic or heterocyclic structure, fused into a pentacyclic D ring, the method includes: Base The reaction agent, the compound of the formula A 2 Shushu: Fe group of 'Compound A 2 1 1 of the formula has the following structure: 0 [A211] wherein ^^-, {^, ^^, ruler, and ^-^ have the same meanings as defined above, and R17 is (^ to (: 4 alkyl groups). 16. A method for preparing compounds conforming to formula IE Method: 〇 Of which -eight-eight-represents a group-〇11114-0: 11115- or &lt; 114 = 〇115- -12- This paper size applies the Chinese National Standard (CNS) A4 specification (210 X 297 mm) 8 8 8 8 A BCD VI. Patent application Fanyuan R3, R4 and R5 are selected from the group consisting of fluorene, 4 primes, ^ ^ ^ ^ ^ ^, hydroxy, low carbon number, low carbon number, alkyl group, hydroxyalkyl group, environmental group , Aryloxy, ... group, carbonyl group, # R26 is c 1 to c4 alkyl group;-BB- represents the group -CHR6_CHR70 and the α- or cold-positioned group: R6 • C Η · —— C Η-III wherein R6 and R7 are respectively selected from the group consisting of hydrogen, halogen, low-carbon alkoxy, fluorenyl, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxycarboxy, alkoxyalkyl, and cyano This method includes: alkylating a compound of formula 2 丨 by reacting with an alkylating agent in the presence of a base, and the compound of formula 2 1 1 has the following structure: [211] Of which-A- A-, R3 and-B- B- are as good as above. 17. —A method for preparing a compound conforming to formula 2 1 1: 13- This paper size applies to Chinese National Standard (CNS) A4 (210 X 297 mm) 593305 A8 B8 C8 D8 6. Scope of patent application -Eight-eight-representing group _chr4-chr5 · or _cr4 = cr5 · R, R and R5 are selected from the group consisting of hydrogen, halogen, guanyl, low carbon number alkynyl, low number alkynyl, hydroxyalkyl , Alkoxyalkyl, hydroxycarbonyl, cyano, aryloxy, -3-6- represents the group _ (: 111 ^ (:) ^ 7_ or (: 1: _ or / 3-azimuth group: R6 C Η — C Η 2 — R7 CH IU where R6 and R7 are respectively selected from the group consisting of hydrogen, halo, low-carbon alkoxy, fluorenyl, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, and alkoxy Carbonyl, alkoxyalkyl, cyano, aryloxy, R8 / and R9G are respectively selected from R8 * R9, or chiral 80 and r9G together to form a ketone. R and R are respectively selected from the group consisting of hydrogen, dentin, and lower alkane Oxy, arsenite, k-based, k-oxyalkyl, light carbonyl, oxo, oxocarbonyl, oxoalkyl, cyano, aryloxy, or R8 and R9 together contain a carbocyclic or heterocyclic The structure of the ring, or R8 or R9 and R6 or R7-from the contract -14- GM China National Standard τ 593305 A8 B8 C8 D8 contains a mosquito ring or heterocyclic structure and is fused into a five-ring D ring. The method includes: oxidizing a compound of formula 2 10 &lt; the compound of formula 2 10 has the following structure: [A210] wherein -A-A-, -B_B-, R3, R8G, and R90 are as defined above. 18. The method according to item 17 of the scope of patent application, wherein R8 and R9 include X representing two hydrogen atoms, oxo or = s, Y1 and Y2 together represent an oxygen bridge _〇-, or Y1 represents a hydroxyl group, and Y2 Represents a mesityl group, a lower carbon number alkyl group, or if X represents fluorene], it may also represent an acid oxy group. 19. Method according to item 18 of the scope of patent application, wherein R8 and R9 include 20 ’methods for preparing compounds [A211] where -AA- represents a group-CHR4-CHR5-* _ CR4 = CR5-R, R and R are selected from the group consisting of hydrogen, halogen, meridian, low-carbon alkyl, low-sulphonic plate oxygen, per-alkyl group , Oxyalkyl, carboxyl, cyano, aryloxy, θ -BB- represents the group -CHR6-CHR7- or α · or / 5-azimuth group: R6-· -C Η -C Η 2 -C Η 一 III in which R6 and R7 are respectively selected from the group consisting of hydrogen, quotient, low-carbon alkoxy, fluorenyl, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxygenic group, and B An oxo group, an oxo group, and an aryloxy group, and R8 and R9 are respectively selected from the group consisting of hydrogen, halogen, alkoxy, fluorenyl, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, Alkoxycarbonyl, alkoxyalkyl, cyano, aryloxy, or a structure in which R8 and R9 together contain a carbocyclic or heterocyclic ring, or a rule of 8 or 119 and 116 or 117 together Applicable to China National Standard (CNS) A4 specification (210 X 297 public directors) 593305 Containing a carbocyclic or heterocyclic structure, the method includes: fused into a five-ring D ring, and reacting the formula A 2 0 9-3 __ with _ $, the formula A209C, the 7-hemal acetal intermediate and peroxidation The oxygen compound conforms to the following formula: [A209C] wherein-r &gt; 8 -f ^ Q, R, R, and R are as defined above. 21. According to the patent application Fan! The method of yoke 20 items, wherein the R8 and R9 include / CV7), M ,, (° Η2) 2. C-XX represents two hydrogen atoms, oxo or = s, Y and Y2 together represent oxygen The bridge ·-, or γ 1 represents a #i group, and Y 2 represents a hydroxyl group, a lower carbon number alkyl group, or if χ represents h 2, an alkylalkoxy group may also be substituted. 22. The method according to item 21 of the patent application, wherein the R8 and R9 include 0 Ί -17-^ Zhang standard 11 ^ Home Standard (CNS) A4 specifications (View 297 public love) &quot; ~: --- -~. 593305 8 8 8 8 A BCD patent application scope 23 · —A method for preparing a compound conforming to the following formula 〇—— OH [A210] where -AA- represents a group-CHR4-CHR5 · or -CR4 = CR5-R3, R4 and R5 are selected from the group consisting of hydrogen, halogen, hydroxyl, low-carbon alkyl, and low-carbon alkoxy , Hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, cyano, aryloxy, -BB- represents the group -CHR6-CHR7- or [or / 3-azimuth group: R6 • CH — CH,- CH IU where R6 and R7 are selected from the group consisting of hydrogen, halogen, low-carbon alkoxy, fluorenyl, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxycarbonyl, alkoxyalkyl, and cyano And aryloxy, and R8G and R9G are respectively selected from the group consisting of r9, or ruler 80 and ruler 9 () to form a ketone. R and R are respectively selected from the group consisting of hydrogen, tooth element, low carbon number alkoxy group, brewer group, and Alkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxycarbonyl-18-This paper size applies to China National Standard (CNS) A4 (210 X 297 mm) 593305 A8 B8 C8 , Donkey oxyalkyl, cyano, aryloxy, or R8 and R9 together contain a mosquito frame or heterocyclic structure, or R8 or R9 and R6 or R7 together contain a slave% or heterocyclic structure, Fusion into a five-ring D ring, the method includes: reacting a 3-keto 5,7-hemiacetal intermediate of formula A 2 0 9 C with a peroxide oxidant. The compound of formula A209C conforms to the following formula: [A209C] wherein -A · A_, -B-B-, R3, R8, and R9 are as defined above. 24. The method according to item 23 of the scope of patent application, wherein the R8 and R9 include 7) 丨 丨 丨 丨 丨 (CH2) 2 · X represents two hydrogen atoms, oxo or = S, Y1 and Y2 together represent oxygen The bridge _〇-, or γ 1 represents a hydroxyl group, and Y 2 represents a hydroxyl group, a lower carbon number alkyl group, or if X represents h 2, it may also represent an alkoxy group. 25. The method according to item 24 of the scope of patent application, wherein the R8 and R9 include -19- this paper size applies the Chinese National Standard (CNS) A4 specification (21 × x7 mm) 593305 8 8 8 8 A BCD application Patent scope 0 II 26.-A method for preparing a compound according to the following formula R90 [A209] -AA- represents a group-CHR, CHR5- or -CR4 = CR5- R3, R4 and R5 are selected from the group consisting of hydrogen, halogen, hydroxyl, low-carbon alkane J, low-carbon alkoxy, hydroxyalkane , Alkoxyalkyl, hydroxycarbonyl, rudi, aryloxy, -BB · represents a group -CHR6-CHR7- or α- or / 5-azimuth group, R6 -C Η — C Η 2 — C Η-where R6 and R7 are selected from the group consisting of hydrogen, halogen, low-carbon alkoxy ', fluorenyl, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, alkyl, alkanoyl, alkoxyalkyl, cyano Group, aryloxy group, and R80 and R9G are respectively selected from the group of 8 and 1, * r8 (^ r9G together form a ketone. R8 and R9 are respectively selected from the group consisting of hydrogen, halogen, low-carbon alkoxy group, commercial group, hydroxyalkyl , Alkoxyalkyl, Hydroxycarbonyl, Alkyl, Alkoxycarbonyl 3 -20-This paper is suitable for wealth and money (CNH ^ 格 (⑽x 297 公 董) 593305 8 8 8 8 AB c D Oxyfe, chloro, aryloxy, or R8 and R9 together contain a carbocyclic or heterocyclic structure, or R8 or R9 and R6 or R7 contain a carbocyclic or heterocyclic structure, fused into D ring of five rings, and -Ε · Ε · is selected from the group consisting of: \ 1 X LII: XLIV OR · I, c—2; XLV and Ro—c R ίο ——R XL VI ο XLVII .21 C—CR -21-This paper size applies to China National Standard (CNS) A4 specification (210X 297 Mm) 593305 A8,. B8 C8 D8 6. The scope of patent applications R21, R22 and R23 are selected from hydrogen, alkyl, halo, nitro and cyano, respectively, and R24 is selected from the group consisting of hydrogen and low-carbon alkyl, The method includes: hydrolyzing a compound conforming to formula A208 [A208] wherein _Α_Α_, _Β · Β_, · Ε_Ε_, R3, R80, and R90 have the same meanings as defined above, and the alkyl or R19 0 · groups together form a 0,0-oxyl-extension group bridge; and an alkyl group. 27. A method for preparing a compound according to the following formula: [A205] where -AA- represents the group-CHR4-CHR5- or -CR4 = CR5- -22- This paper size applies to China National Standard (CNS) A4 specifications (210 X 297 mm) 593305 8 8 8 8 A BCD R3, R4, and R5 are selected from the group consisting of hydrogen, halo, hydroxy, lower alkyl, lower alkoxy, light alkyl, alkyl, hydroxycarbonyl, cyano, and aryloxy Group: R6 -C Η — C Η 2 ~ 丨 _ C Η-III, where R and R7 are selected and selected from the group consisting of hydrogen, functional element, low carbon number alkoxy group, 酉 盛 基 ―, mesomorphic group, and morphological oxygen An alkyl group, an alkyl group, an alkyl group, an alkyloxycarbonyl group, an alkyloxyalkyl group, a cyano group, an aryloxy group, and an alkyl group or an R19 0-group together form a 0,0-oxyalkylene bridge; and an alkyl group. Where -E-E- is selected from the group consisting of: XLIH XLIV R--c // RI 2 ocR VLXR 〇——c R. I c ——R XLVI 23 This paper size applies to China National Standard (CNS) A4 (210 x 297 mm) 593305 8 8 8 8 A BCD ^, the scope of application for patents and 〇2, XLVII R21, R22 and R23 are respectively selected from hydrogen, alkyl, autogen, nitro and cyano, and R24 is selected from hydrogen and low-carbon alkyl. The method includes: The compound of formula 204 reacts with a lower carbon number of alcohols and acids. The compound of formula 204 has the following structure: 9 OH [A204] where _A-A_, -B-B_, -E-E-, R3 and R19 are as defined above. 28 ·-A method for preparing a compound according to the formula: [A204] Among which -24 paper sizes are applicable to Chinese National Standard (CMS) A4 specifications (210 x 297 mm) 593305 8 8 8 8 A BCD patent application scope -AA- representative group-CHR4-CHR5- or -CR4 = CR5- R3, R4 and R5 are selected from the group consisting of hydrogen, halogen, hydroxy, low-carbon alkyl, low-carbon alkoxy, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, cyano, and aryloxy,- BB- represents a group of ΟΗΙ16 · (: ΗΙ17_ or α- or cold-azimuth group: R6 III wherein R6 and R7 are respectively selected from the group consisting of hydrogen, halogen, low-carbon alkoxy, fluorenyl, and hydroxyalkyl , Alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxycarbonyl, alkoxyalkyl, cyano, and aryloxy, and the 19 is a CrCU alkyl or R19 0-group together to form 0,0-oxyalkylene Base bridge; where -EE- is selected from the following: .22 \ 1 XL: I C- XL IV OR2 Call XLV RO 1C R nc IR 25 XI This paper size applies to China National Standard (CNS) A4 (210 X 297 mm) ) 593305 8 8 8 8 AB c D patent application scope and XLVII R21, R22 and R23 are respectively selected from hydrogen, alkyl, halogen, nitro and cyano, and R24 is selected from hydrogen and low-carbon alkyl, The method comprises: hydrolyzing a compound conforming to the formula 230, the compound having the following structure: [A203] in which-A- A-, -BB-, -EE-, R3 and R18 are as defined above and the rule "is a d-alkyl group or a R18 group together to form a 〇-〇-oxyalkylene基 桥 〇29. — A method for preparing a compound according to the following formula: [A204] -26- This paper size applies to Chinese National Standard (CNS) A4 specification (210X 297 mm) 593305 8 8 8 8 AB c D 々, the scope of patent application where -AA- represents the group _ (: 11114- ( : ^ 1115- or -CR4 = CR5- R3, R4 and R5 are selected from the group consisting of hydrogen, halogen, hydroxyl, low-carbon alkyl, low-carbon alkoxy, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, cyanide Group, aryloxy group, -BB- represents a group -CHR6-CHR7- or α- or / 3-azimuth group: R6 Wherein R6 and R7 are respectively selected from the group consisting of hydrogen, halogen, low-carbon alkoxy, fluorenyl, lightly burned, oxyhalo, alkynyl, alkynyl, alkoxycarbonyl, alkoxyalkyl, cyano, and aromatic. Oxygen, and -EE- is selected from the group consisting of: R21 R22 \ 1 丨 CC—— /) p23 XLIII XLIV -27- This paper size is in accordance with Chinese National Standard (CNS) A4 (210 X 297 mm) 593305 8 8 8 8 ABCD VI. Patent application scope OR \ 1 and XLVI XLVII R18 are C ^ C: 4 alkyl, or .Ru〇_ group together to form a 〇, 〇_ oxy alkynyl bridge 'R21, respectively selected from hydrogen, alkynyl, halogen, synyl and cyano, R2 · 4 is selected from hydrogen and lower carbon alkyl The method comprises: protecting the keto substituent of the compound of the formula A2O1 by reacting with the chain alcohol and the reactant in the presence of an orthoformate under acidic conditions, the compound of the formula having the following structure: [A201] From this, -AA-, -BB-, _E_E_ * R3 and as defined above are used to produce a 3-dilute alkyd intermediate that conforms to the formula 2 0 2 · 28 This paper applies the Chinese National Standard (CNS) A4 Specifications (210 X 297 mm) -------- 593305 A8 B8 C8 D8 [A202] wherein -eight-eight-,-; 8-; 6-,-£ -ugly-, ruler 3 are as defined above, and 1118 is a C, C4 alkyl or R18 O group. Together, it forms 0, 0-alkylene bridge; and reducing the compound of formula 202 to produce a compound conforming to formula A203: [Α203] wherein -Α-Α-, -Β-Β-, -Ε-Ε-, R3 and R18 are as defined above; and a compound conforming to formula A203 is hydrolyzed to thereby produce a compound of formula A204. 30. A method for preparing a compound according to the following formula: CHO [A203] -29- This paper size applies to Chinese National Standard (CNS) A4 specification (210 X 297 mm) 593305 8 8 8 8 A BCD patent application scope where -AA- represents the group-CHR4-CHR5- or- CR4 = CR5- R3, R4 and R5 are selected from the group consisting of hydrogen, halo, hydroxyl, low-carbon alkyl, low-carbon alkoxy, hydroxyalkyl, alkoxyalkyl, hydroxycarbonyl, cyano, and aryloxy , -BB- represents a group -CHR6-CHR7- or α- or / 3-azimuth group: R6 III wherein R6 and R7 are respectively selected from the group consisting of hydrogen, halogen, low-carbon alkoxy, fluorenyl, and hydroxyalkane Group, alkoxyalkyl, hydroxycarbonyl, alkyl, alkoxycarbonyl, alkoxyalkyl, cyano, aryloxy, and -E_Ε- is selected from the group consisting of: 22 \ 1 XLI: XLIV XLV XLVI -30- This paper size applies to Chinese National Standard (CNS) A4 (210 x 297 mm) 593305 A8 B8 C8 D8 VI. Patent application scope and CR21 XLVII R18 is alkyl, or C-17 The R18〇- groups at the same position together form a 0,0-oxyalkylene bridge, R21, R22 and R23 are respectively selected from hydrogen, alkyl, halogen, nitro and cyano, and R24 is selected from hydrogen and low carbon number The method includes: reducing a compound conforming to Formula A202. [A202] wherein -A-A-, -B-B-, -E-E-, R3 and R18 are as defined above. 31. The method according to any one of claims 9 to 30 in the scope of the patent application, wherein r3 is hydrogen, -A-A- represents a group -CHR4-CHR5_, and _Β · Β · represents a group -CHR6-CHR7. 32. The method according to item 31 of the scope of patent application, wherein R4, r5, ruler 6 and ruler 7 are each hydrogen. Method, in which R3′_B-B • represents the base figure 33. According to any one of the 26th to 30th scope of the patent application, it is hydrogen, and -AA- represents the group -CHR4-CHR5--CHR6-CHR7, -31- This paper size applies to Chinese National Standard (CNS) A4 specifications (210 X 297 mm) 593305 A8 B8 C8 D8 Six, the scope of patent applications R4, R5, R6 and R7 are each hydrogen, and -E · Ε- is selected from the following and XLIII XLVII, nitro and R21, R22 'and R23 are selected from hydrogen, alkyl and halocyano, respectively. 34.-a compound of formula D, 32 -32- This paper size applies to Chinese National Standard (CNS) A4 (210 X 297 mm) 593305 A8 B8 This paper size is in accordance with Chinese National Standard (CNS) A4 (210X 297 mm) 593305 8 8 8 8 A B c D 6. Scope of patent application 0 38.—Compounds of formula F, ο 4 2 Wherein -A-A- represents the group -CH2-CH2 · or -CH = CH-. 39. The compound according to item 38 of the application, which has the following formula: Physicochemical -34- The paper scale is applicable to Chinese National Standard (CNS) A4 specifications (210X 297 mm) 593305 8 8 8 8 A BCD 々 Scope of patent application Where -A-A- represents the group -CH2-CH2- or -CH = CH-; and R80 and R90 are each a hydroxyl group or a hydroxycarbonylalkyl group or form a ketone or represent the following groups. 41. The compound according to item 40 of the scope of patent application, which has the formula 〇 42.—A compound of formula 2 10, -35- This paper size applies Chinese National Standard (CNS) A4 specification (210 X 297 mm) 593305 8 8 8 8 AB c D The scope of patent application where -AA- represents the group -CH2-CH2- or -CH = CH_ And R8G and R9G are each a hydroxyl group or a hydroxycarbonyl alkyl group or form a ketone or represent the following group. 43. The compound according to item 42 of the scope of patent application, which has the following formula 44. A compound of formula 209 -36- This paper size applies Chinese National Standard (CNS) A4 specification (210 X 297 mm) 593305 8 8 8 8 AB c D. The scope of patent application is -AA- represents the group -CH2_CH2- or -CH = CH-; And R8G and R90 are each a hydroxyl group or a hydroxycarbonyl alkyl group or together form a ketone or represent the following groups. C (17) and -E-E- are selected from the group JCM——CHj- XLIII.〉 C XLIV, OH, I: CH—CH- XLV, OH XLVI, and &gt; Δη- XLVII45 · According to the patent application The 44th scope compound has the following formula: 37- This paper size applies to Chinese National Standard (CNS) A4 (210 X 297 mm) 593305 A8 B8 C8 This paper size applies to China National Standard (CNS) A4 (210 x 297 mm) 593305 A8 B8 C8 D8 Patent application scope &gt; Η-CH2 ^ C = CH- OH, I CH-CH-XL-IV, XLV, OH -CH, XLVI. And A? CH- XLVII 〇 47. According to the scope of application of the 46th patent compound, which has the following formula Hac 48.—Compound of Formula 207, -39-This paper size applies to Chinese National Standard (CNS) A4 (210 X 297 mm) 593305 8 8 8 8 A 207 where -AA- represents the group -CH2-CH2- or -CH = CH-; R19 is an alkyl group or a 190-group-group to form a 0,0-oxoalkylene bridge; R2G is a CVC4 alkyl group; R25 is (^ To alkylene "and -EE- are selected from the following groups: CH --- CH :, XLIII, and &gt; = CH-OH> H-L OH ^ X-CH2 &gt; Δη- XLIV, XLV, XLVI, XL VII -40- This paper size applies to Chinese National Standard (CNS) A4 (210 x 297 mm) 593305 8 8 8 8 AB c D Application scope of patent 49. According to the 48th compound of the scope of patent application, It has the following formula. 50.- Compound of Formula 206 206 where -AA- represents the group -CH2-CH2- or -CH = CH-; R19 is (^ to alkyl or R190- groups together to form a 0,0-oxyalkylene bridge; alkyl; and -EE -It is selected from the following groups: XLIII, -CH2; C = CH- XLIV, -41 This paper size is applicable to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) 593305 8 8 8 8 A BCD VI. Application Patent scope and OH, I CH ._-. CH · XLV, OH-CH2 XLVI, XLVII ο 5 51. The compound according to the item in the scope of patent application, which has the following formula H3CO-HjCO ' 〇CHa 52.—a compound of the formula 205 205 42 This paper size applies Chinese National Standard (CNS) A4 specification (210 X 297 mm) 593305 8 8 8 8 A BCD patent application scope where -AA- represents the group -CH2-CH2- or -CH = CH-; R19 is an alkyl group or R190- group that together forms a 0,0-oxoalkylene bridge; an alkyl group; and -EE- is selected from the group: _CH, XLIII, = CH- XLIV, OH, I / CH—CH_ • XLV, OH di 0-Cha * XLVI, and Λ feH- XLVII 53. The compound according to item 52 of the scope of patent application, which has the following formula -43 This paper size is applicable to Chinese National Standard (CNS) A4 specification (210 X 297 mm) 593305 8 8 8 8 A BCD 々, patent application scope 54.— compound of formula 2 04, 4 ο 2 where -AA- represents the group -CH2-CH2- or _CH = CH-; R19 is CiSC4 alkyl or R190- group-groups together to form a 0,0-oxoalkylene bridge; and -EE- system selection From the following groups Η —-C H2-XLIII,: CH_ XLIV, \ I CH * —Ch ..... XLV, OH X-ch2_ XLVI, and Xinggong 44 This paper applies the Chinese National Standard (CNS) A4 Specifications (210 X 297 mm) 593305 8 8 8 8 A BCD VI. Patent Application Range 5 5. The compound according to item 54 of the patent application range has the following formula 56.—A compound of formula 203, R1 8 R1 3 ο 2 where -AA- represents the group -CH2-CH2- or -CH = CH-; R18 is (^ to alkyl or C-17 or R180- groups together to form 0,0-oxygen Alkylene bridge; and -EE- is selected from the following groups: / rCH-CH2- XLIII, &gt;: CH · XLIV, -45- This paper size applies to China National Standard (CNS) A4 (210 X 297 mm) ) 593305 8 8 8 8 AB c D Patent application park OH XLV, ^ CH-CH- Oh-ch2- XLVI (and XLVII 57. According to the 56th patent application, it has the following formula -46 This paper size applies to China National Standard (CNS) A4 (210 X 297 mm)