HK1234744B - Antifungal compound process - Google Patents

Description

Method for preparing antifungal compound

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Patent Application No. 61/955,615, filed March 19, 2014, which is expressly incorporated herein by reference.

The present invention relates to a method for preparing compound 1 which is useful as an antifungal agent. In particular, the present invention aims to provide a novel method for preparing compound 1 and its substituted derivatives.

Government Support Statement

This invention was made under a cooperative research and development agreement with the National Institutes of Health, an agency of the Department of Health and Human Services. The U.S. Government has certain rights in this invention.

Background Art

Living organisms have developed a rigorous process for specifically importing metals, transporting them to intracellular storage sites, and ultimately transporting them to useful locations. One of the most important functions of metals such as zinc and iron in biological systems is to enable the activity of metalloenzymes. Metalloenzymes are enzymes that bind metal ions to the enzyme's active site and utilize the metal as part of a catalytic process. More than one-third of all characterized enzymes are metalloenzymes.

The function of metalloenzymes is highly dependent on the presence of metal ions in the active site of the enzyme. It is well recognized that agents that bind to and inactivate the active site metal ions significantly reduce the activity of the enzyme. Nature uses this same strategy to reduce the activity of certain metalloenzymes during periods when enzymatic activity is not required. For example, the protein TIMP (tissue inhibitor of metalloproteinase) binds zinc ions in the active site of various matrix metalloproteinases, thereby preventing enzymatic activity. The pharmaceutical industry has used the same strategy in the design of therapeutic agents. For example, the azole antifungals fluconazole and voriconazole contain a 1-(1,2,4-triazole) group that binds to the heme iron present in the active site of the target enzyme lanosterol demethylase, thereby inactivating the enzyme.

In the design of clinically safe and effective metalloenzyme inhibitors, the use of the most suitable metal binding group for a specific target and clinical indication is crucial. If a weakly bound metal binding group is used, the efficacy may be suboptimal. On the other hand, if a very tightly bound metal binding group is used, the selectivity of the target enzyme relative to the related metalloenzyme may be suboptimal. The lack of optimal selectivity can be the cause of clinical toxicity, which is attributed to the unexpected inhibition of these target-external metalloenzymes. An example of this clinical toxicity is the unexpected inhibition of human drug metabolizing enzymes such as CYP2C9, CYP2C19 and CYP3A4 by currently available azole antifungal agents such as fluconazole and voriconazole. It is believed that this target-external inhibition is mainly caused by the indiscriminate binding of the iron in the active site of 1-(1,2,4-triazole) currently used to CYP2C9, CYP2C19 and CYP3A4. Another example is the joint pain that has been observed in many clinical trials of matrix metalloenzyme inhibitors. This toxicity is believed to be related to the inhibition of target-external metalloenzymes, which is attributed to the indiscriminate binding of the hydroxamic acid group to the zinc in the target-external active site.

Therefore, the research of metal binding groups that can achieve better balance of efficacy and selectivity remains an important goal, and is of great significance in the realization of therapeutic agents and methods for meeting the existing unmet needs in treating and preventing its diseases, diseases and its symptoms. Similarly, it is necessary to synthesize the therapeutic agent at laboratory scale and final commercial scale. The synthesis of voriconazole has been achieved by adding metal-based nucleophiles (Zn, Zr, Ce, Ti, Mg, Mn, Li) to azole methyl-substituted ketones (M.Butters, Org.Process Res.Dev.2001, 5, 28-36). In these examples, nucleophiles are ethyl-pyrimidine substrates. Similarly, optically active azole-methyl epoxides are prepared as electrophilic reagent precursors for the synthesis of ravuconazole. (A.Tsuruoka, Chem.Pharm.Bull.1998, 46, 623-630). Nevertheless, the development of methods with improved efficacy and selectivity is needed.

Summary of the Invention

The present invention is directed to a method for synthesizing 1 or 1a. The method may include the compounds described herein. A first aspect of the present invention relates to a method for preparing a compound of formula 1 or a pharmaceutically acceptable salt, hydrate, solvate, complex, or prodrug thereof.

The compounds herein include those in which the compounds are believed to obtain affinity for metalloenzymes at least in part by forming one or more of the following types of chemical interactions or bonds with the metal: sigma bonds, covalent bonds, coordinate-covalent bonds, ionic bonds, pi bonds, delta bonds, or feedback bonding interactions.

Methods for evaluating metal-ligand binding interactions are known in the art, as exemplified in the literature, including, for example, "Principles of Bioinorganic Chemistry", Lippard and Berg, University Science Books, (1994); "Mechanisms of Inorganic Reactions", Basolo and Pearson, John Wiley & Sons Inc; 2nd Edition (September 1967); "Biological Inorganic Chemistry", Ivano Bertini, Harry Gray, Ed Stiefel, Joan Valentine, University Science Books (2007); Xue et al., "Nature Chemical Biology", Vol. 4, No. 2, 107-109 (2008).

In the following aspects, reference is made to the schemes and compounds herein, including reagents and reaction conditions described herein. Other aspects include any compound, reagent, transformation thereof, or method described in the Examples herein (in whole or in part), including embodiments with a single element (e.g., compound or transformation) or embodiments including multiple elements (e.g., compound or transformation).

In one aspect, the present invention provides a method for preparing a compound of formula II:

Involving epoxy ring opening of compounds of formula I:

Obtaining a compound of formula II;

Where R is

In another aspect, the present invention provides a method for preparing amino alcohol 1-6 or 1-7 or a mixture thereof:

This involves arylation of substituted pyridine 4b or 4c or a mixture thereof:

Obtaining compound 1-6 or 1-7 or a mixture thereof;

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl.

In another embodiment, the present invention provides a method for enriching the enantiomeric purity of a mixture of enantiomeric compounds, comprising:

(i) crystallizing a mixture of enantiomeric compounds with a chiral acid in a suitable solvent or solvent mixture wherein:

Suitable solvents or solvent mixtures are selected from acetonitrile, isopropanol, ethanol, water, methanol or a combination thereof;

A mixture of enantiomeric compounds includes

(ii) separating the enantiomerically enriched mixture of compounds

(iii) reslurrying the enantiomerically enriched chiral salt in a slurrying solvent or a mixture of slurrying solvents;

(iv) free-base the enantiomerically enriched chiral salt to provide a mixture of enantiomerically enriched compounds.

In another embodiment, the present invention provides a method for enriching the enantiomeric purity of a mixture of enantiomeric compounds, comprising:

(i) crystallizing a mixture of enantiomeric compounds with a chiral acid in a suitable solvent or solvent mixture wherein:

Suitable solvents or solvent mixtures are selected from acetonitrile, isopropanol, ethanol, water, methanol or a combination thereof;

A mixture of enantiomeric compounds contains

(ii) separating the enantiomerically enriched mixture of compounds;

(iii) free-base the enantiomerically enriched chiral salt to provide a mixture of enantiomerically enriched compounds.

In another aspect, the chiral acid from any of the embodiments provided herein is selected from tartaric acid, dibenzoyltartaric acid, malic acid, camphoric acid, camphorsulfonic acid, ascorbic acid, and di-p-toluoyltartaric acid;

In another aspect, a suitable solvent or solvent mixture from any embodiment provided herein is 1-propanol, 1-butanol, ethyl acetate, tetrahydrofuran, 2-methyltetrahydrofuran, toluene, methyl tert-butyl ether, diethyl ether, dichloromethane, 1,4-dioxane, 1,2-dimethoxyethane, isopropyl acetate, heptane, hexane, cyclohexane, or octane, or a combination thereof.

In another aspect, a suitable pulping solvent or pulping solvent mixture from any embodiment provided herein is 1-propanol, 1-butanol, ethyl acetate, tetrahydrofuran, 2-methyltetrahydrofuran, toluene, methyl tert-butyl ether, diethyl ether, dichloromethane, 1,4-dioxane, 1,2-dimethoxyethane, isopropyl acetate, heptane, hexane, cyclohexane, or octane, or a combination thereof.

In another aspect, a suitable solvent or solvent mixture from any embodiment provided herein is a) acetonitrile or b) a mixture of acetonitrile and isopropanol. Alternatively, in another aspect, a mixture of acetonitrile and methanol comprises 80-90% acetonitrile and 10-20% isopropanol.

In another aspect, a suitable pulping solvent or pulping solvent mixture from any embodiment provided herein is a) acetonitrile or b) a mixture of acetonitrile or isopropanol. Alternatively, in another aspect, a mixture of acetonitrile and isopropanol comprises 80-90% acetonitrile and 10-20% isopropanol.

In another aspect, the present invention provides a method for preparing compound 1 or 1a or a mixture thereof:

This involves converting amide 2c:

Converted into compound 1 or 1a, or a mixture thereof;

wherein R ₁ is halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO _{2 )-substituted alkyl, —O(SO 2 )} -aryl, or —O(SO ₂ )-substituted aryl;

A is N(OMe)Me, NR ₈ R ₉ , or

p is 1, 2, 3, or 4;

q is 1, 2, 3, or 4;

Each of R ₈ and R ₉ is independently H, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, or substituted heteroaryl.

In another aspect, the present invention provides a method for preparing compound 1 or 1a or a mixture thereof:

This involves converting amide 2c:

Converted into compound 1 or 1a or a mixture thereof;

wherein R ₁ is halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO _{2 )-substituted alkyl, —O(SO 2 )} -aryl, or —O(SO ₂ )-substituted aryl;

B is N(OMe)Me, NR ₈ R ₉ , or

X is O, NR ₈ , or S;

r is 2, 3, or 4;

s is 2, 3, or 4;

Each of R ₈ and R ₉ is independently H, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, or substituted heteroaryl.

In another aspect, the present invention provides a method for preparing compound 1 or 1a or a mixture thereof:

comprising morpholine amide 2b:

Converted into compound 1 or 1a or a mixture thereof;

wherein R ₁ is halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO _{2 )-substituted alkyl, —O(SO 2 )} -aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a method comprising subjecting morpholine amide 2b to:

and reactions;

wherein M is Mg or MgX; X is a halogen;

Obtaining compound 1 or 1a, or a mixture thereof;

wherein R ₁ is halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO _{2 )-substituted alkyl, —O(SO 2 )} -aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a method comprising subjecting morpholine amide 2b to:

and

reaction;

wherein M is Mg or MgX, Li, AlX ₂ ; X is halogen, alkyl, or aryl;

Compound 1 or 1a, or a mixture thereof is obtained:

wherein R ₁ is halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO _{2 )-substituted alkyl, —O(SO 2 )} -aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, any of the embodiments provided herein may include amidation of ester 2:

Morpholine amide 2b is obtained:

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, any of the embodiments provided herein may comprise amidation of ester 2d:

Morpholine amide 2b is obtained:

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl;

R ₈ is H, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, or substituted heteroaryl.

In another aspect, any of the embodiments provided herein can include making ester 2:

Reaction with morpholine to give morpholine amide 2b:

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, any of the embodiments provided herein may include:

(i) Replace the morpholinyl group in morpholinamide 2b: to give ketone 3:

(ii) Arylation of ketone 3: Obtaining aryl-pyridine 1-4:

(iii) Formation of aryl-pyridine epoxides 1-4: Obtaining epoxide 5:

(iv) Ring opening of epoxide 5: to give amino alcohol ±1-6:

(v) enriching the enantiomeric purity of the amino alcohol ± 1-6: to obtain enantiomerically enriched amino alcohol 1-6* or 1-7*: or a mixture thereof;

(vi) tetrazolyl to form enantiomerically enriched amino alcohol 1-6* or 1-7*: or a mixture thereof to provide compound 1 or 1a: or a mixture thereof;

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides compound 1-6* or 1-7*:

or a mixture thereof.

In another aspect, the present invention provides a method for preparing enantiomerically enriched arylpyridine 1-6* or 1-7*, enantiomerically enriched amino alcohol 1-6* or 1-7*: or a mixture thereof, the method comprising:

(i) Replace the morpholinyl group in morpholinamide 2b: to give ketone 3:

(ii) Arylation of ketone 3: Obtaining aryl-pyridine 1-4:

(iii) Formation of the epoxide of aryl-pyridine 1-4: to give epoxide 5:

(iv) Ring opening of epoxide 5: to give amino alcohol ±1-6:

(v) enriching the enantiomeric purity of the amino alcohol ± 1-6: to obtain enantiomerically enriched amino alcohol 1-6* or 1-7*: or a mixture thereof;

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO _{2 )-substituted alkyl, —O(SO 2 )} -aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a method for preparing enantiomerically enriched arylpyridine 1-6* or 1-7*, enantiomerically enriched amino alcohol 1-6* or 1-7*: or a mixture thereof, the method comprising:

(i) Ring opening of epoxide 5: to give amino alcohol ±1-6:

(ii) enriching the enantiomeric purity of the amino alcohol ± 1-6: to obtain enantiomerically enriched amino alcohol 1-6* or 1-7*: or a mixture thereof;

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a method for preparing epoxide 5, comprising:

(i) Replace the morpholinyl group in morpholinamide 2b: to give ketone 3:

(ii) Arylation of ketone 3: Obtaining aryl-pyridine 1-4:

(iii) Formation of the epoxide of aryl-pyridine 1-4: to give epoxide 5:

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO _{2 )-substituted alkyl, —O(SO 2 )} -aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a method for preparing epoxide 5:, the method comprising:

(i) Formation of the epoxide of aryl-pyridine 1-4: to give epoxide 5:

In another aspect, any of the embodiments provided herein may include:

(i) Replace the morpholinyl group in morpholinamide 2b: to give ketone 3:

(ii) Formation of the epoxide of ketone 3: to give epoxide 4:

(iii) Ring opening of epoxide 4: gives amino alcohol ± 4b:

(iv) Enantiomeric purity of the enriched amino alcohol ± 4b: to obtain enantiomerically enriched amino alcohol 4b or 4c:

or a mixture thereof;

(v) tetrazolyl esters that form enantiomerically enriched amino alcohols 4b or 4c: or mixtures thereof, to afford tetrazolyl esters 6 or 6a: or mixtures thereof;

(vi) arylating tetrazole 6 or 6a: or a mixture thereof to obtain compound 1 or 1a: or a mixture thereof;

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, any of the embodiments provided herein may include:

(i) Replace the morpholinyl group in morpholinamide 2b: to give ketone 3:

(ii) Formation of the epoxide of ketone 3: to give epoxide 4:

(iii) Ring opening of epoxide 4: gives amino alcohol ± 4b:

(iv) Enantiomeric purity of the enriched amino alcohol ± 4b: to obtain enantiomerically enriched amino alcohol 4b or 4c:

or a mixture thereof;

(v) arylating the enantiomerically enriched amino alcohol 4b or 4c: or a mixture thereof to provide the enantiomerically enriched aryl pyridine 1-6* or 1-7*: or a mixture thereof;

(vi) generating enantiomerically enriched arylpyridine 1-6* or 1-7*: or a mixture thereof to obtain compound 1 or 1a: or a mixture thereof;

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a method for preparing enantiomerically enriched arylpyridine 1-6* or 1-7*, enantiomerically enriched amino alcohol 1-6* or 1-7*: or a mixture thereof, the method comprising:

(i) Replace the morpholinyl group in morpholinamide 2b: to give ketone 3:

(ii) Formation of the epoxide of ketone 3: to give epoxide 4:

(iii) Ring opening of epoxide 4: to provide amino alcohol ± 4b:

(iv) Enantiomeric purity of the enriched amino alcohol ± 4b: to obtain enantiomerically enriched amino alcohol 4b or 4c:

or a mixture thereof;

(v) arylating the enantiomerically enriched amino alcohol 4b or 4c: or a mixture thereof to provide the enantiomerically enriched aryl pyridine 1-6* or 1-7*: or a mixture thereof;

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a method for preparing enantiomerically enriched arylpyridine 1-6* or 1-7*, enantiomerically enriched amino alcohol 1-6* or 1-7*: or a mixture thereof, the method comprising:

(i) Ring opening of epoxide 4: to give amino alcohol ± 4b:

(ii) enriching the enantiomeric purity of the amino alcohol ± 4b: to obtain enantiomerically enriched amino alcohol 4b or 4c: or a mixture thereof;

(iii) arylating the enantiomerically enriched amino alcohol 4b or 4c: or a mixture thereof to obtain the enantiomerically enriched aryl pyridine 1-6* or 1-7*: or a mixture thereof;

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO _{2 )-substituted alkyl, —O(SO 2 )} -aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a method for preparing enantiomerically enriched amino alcohol 4b or 4c: or a mixture thereof, the method comprising:

(i) Replacement of the morpholinyl moiety in morpholinamide 2b: to give ketone 3:

(ii) Formation of the epoxide of ketone 3: to give epoxide 4:

(iii) Ring opening of epoxide 4: gives amino alcohol ± 4b:

(iv) enriching the enantiomeric purity of the amino alcohol ± 4b: to obtain enantiomerically enriched amino alcohol 4b or 4c: or a mixture thereof;

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl;

In another aspect, the present invention provides a method for preparing enantiomerically enriched amino alcohol 4b or 4c: or a mixture thereof, the method comprising:

(i) Ring opening of epoxide 4: to give amino alcohol ± 4b:

(ii) Enantiomeric purity of the enriched amino alcohol ± 4b: to obtain enantiomerically enriched amino alcohol 4b or 4c:

or a mixture thereof;

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a method for preparing enantiomerically enriched amino alcohol 4b or 4c: or a mixture thereof, the method comprising:

(i) Enantiomeric purity of the amino alcohol ± 4b: is enriched to obtain enantiomerically enriched amino alcohol 4b or 4c:

or a mixture thereof;

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a method for preparing compound 1 or 1a or a mixture thereof:

Involving epoxy ring opening of compounds of Formula I, VII or VIIa:

To obtain a compound of formula II, VIII or VIIIa:

wherein each R ₂ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a method for preparing compound 1 or 1a or a mixture thereof:

This involves the formation of substituted pyridines 4d or 4e or mixtures thereof:

tetrazole to form 6c or 6d: or a mixture thereof;

wherein each R ₂ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a method for preparing compound 1 or 1a or a mixture thereof:

This involves arylating amino alcohol 4b or 4c: or a mixture thereof to provide aryl-pyridine 1-6* or 1-7*: or a mixture thereof,

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a method for preparing compound 1 or 1a or a mixture thereof:

This comprises converting a compound of formula 15 into compound 1 or 1a:

wherein R ₁ is halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO _{2 )-substituted alkyl, —O(SO 2 )} -aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a process for preparing a compound of formula IX or IXa or a mixture thereof:

wherein each Z is independently aryl, substituted aryl, alkyl, or substituted alkyl.

In another aspect, the present invention provides a compound of formula XI or XIa, or a mixture thereof:

in:

each R ₁₀ is independently H, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl, or optionally substituted heteroarylalkyl;

each R ₁₁ is independently H, OH, optionally substituted alkyl, optionally substituted alkoxy, or OC(O)R ₁₄ ;

each R ₁₂ is independently H, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl, or optionally substituted heteroarylalkyl;

R ₁₃ is independently H, OH, optionally substituted alkyl, optionally substituted alkoxy, or OC(O)R ₁₄ ;

each R ₁₄ is independently H, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl, or optionally substituted heteroarylalkyl;

Each t is independently 0, 1, 2 or 3.

In another aspect, R ₁₀ is H and t is 1.

In another aspect, R ₁₂ is H and t is 1.

In another aspect, R ₁₀ is H, R ₁₂ is H, and t is 1.

In another aspect, R ₁₁ is OH or OC(O)R ₁₄ (preferably, OC(O)-p-tolyl), and t is 1.

In another aspect, R ₁₃ is OH or OC(O)R ₁₄ (preferably, OC(O)-p-tolyl), and t is 1.

In another aspect, R ₁₁ is OH or OC(O)R ₁₄ (preferably OC(O)-p-tolyl), R ₁₃ is OH or OC(O)R ₁₄ (preferably OC(O)-p-tolyl), and t is 1.

In another aspect, R ₁₀ is H, R ₁₁ is OH or OC(O)R ₁₄ (preferably OC(O)-p-tolyl), R ₁₂ is H, R ₁₃ is H, OH, or OC(O)R ₁₄ (preferably OC(O)-p-tolyl), and t is 1.

In another aspect, R ₁₀ is H, R ₁₁ is OH or OC(O)R ₁₄ (preferably OC(O)-p-tolyl), R ₁₂ is H, R ₁₃ is OH or OC(O)R ₁₄ (preferably OC(O)-p-tolyl), and t is 1.

In another aspect, R ₁₀ is H, R ₁₁ is OC(O)R ₁₄ (preferably OC(O)-p-tolyl), R ₁₂ is H, R ₁₃ is OC(O)R ₁₄ (preferably OC(O)-p-tolyl), and t is 1.

In another aspect, R ₁₀ is H, R ₁₁ is OC(O)R ₁₄ , R ₁₂ is H, R ₁₃ is OC(O)R ₁₄ , each R ₁₄ is independently optionally substituted arylalkyl, and t is 1. In another aspect, each R ₁₄ is p-tolyl.

In another aspect, R ₁₁ is OH, R ₁₃ is H, and t is 1.

In another aspect, R ₁₀ is H, R ₁₁ is OH, R ₁₂ is H, R ₁₃ is H, and t is 1.

In another aspect, the present invention provides a process for preparing a compound of formula IX or IXa or a mixture thereof:

(i) combining compound 1 or 1a: or a mixture thereof, a sulfonic acid, and a crystallization solvent or a crystallization solvent mixture;

(ii) diluting the mixture from step (i) with a crystallization co-solvent or a mixture of crystallization co-solvents;

(iii) isolating the compound of Formula IX or IXa or a mixture thereof;

wherein each Z is independently aryl, substituted aryl, alkyl, or substituted alkyl.

In another aspect, the present invention provides compound X or Xa or a mixture thereof:

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a method for preparing compound 1 or 1a or a mixture thereof, comprising:

(i) Replace the ester group in ester 2 to obtain ketone 3:

(ii) Formation of the epoxide of ketone 3: to give epoxide 4:

(iii) Ring opening of epoxide 4: gives amino alcohol ± 4b:

(iv) Enantiomeric purity of the enriched amino alcohol ± 4b: to obtain enantiomerically enriched amino alcohol 4b or 4c:

or a mixture thereof;

(v) Producing enantiomerically enriched amino alcohol 4b or 4c:

or a mixture thereof of tetrazoles to produce tetrazoles 6 or 6a: or a mixture thereof;

(vi) arylating tetrazole 6 or 6a: or a mixture thereof to obtain compound 1 or 1a: or a mixture thereof;

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a method for preparing compound 1 or 1a or a mixture thereof, comprising:

(i) Replace the ester group in ester 2 to obtain ketone 3:

(ii) Formation of the epoxide of ketone 3: to give epoxide 4:

(iii) Ring opening of epoxide 4: gives amino alcohol ± 4b:

(iv) Enantiomeric purity of the enriched amino alcohol ± 4b: to obtain enantiomerically enriched amino alcohol 4b or 4c:

or a mixture thereof;

(v) arylating the enantiomerically enriched amino alcohol 4b or 4c: or a mixture thereof to provide the enantiomerically enriched amino alcohol 1-6* or 1-7*: or a mixture thereof;

(vi) tetrazolyl to form enantiomerically enriched amino alcohol 1-6* or 1-7*: or a mixture thereof to provide compound 1 or 1a: or a mixture thereof;

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl.

In another aspect, the present invention provides a method for preparing compound 1 or 1a or a mixture thereof, comprising:

(i) Displace the ester group of ester 2: to give ketone 3:

(ii) Formation of the epoxide of ketone 3: to give epoxide 4:

(iii) Ring opening of epoxide 4: gives amino alcohol ± 4b:

(iv) Enantiomeric purity of the enriched amino alcohol ± 4b: to obtain enantiomerically enriched amino alcohol 4b or 4c:

or a mixture thereof;

(v) arylating the enantiomerically enriched amino alcohol 4b or 4c: or a mixture thereof to provide the enantiomerically enriched amino alcohol 1-6* or 1-7*: or a mixture thereof;

(vi) forming an enantiomerically enriched salt of amino alcohol 1-6* or 1-7*: or a mixture thereof to obtain compound XI or Xia: or a mixture thereof;

(vii) generating a tetrazole of XI or Xia: or a mixture thereof to obtain compound 1 or 1a: or a mixture thereof;

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl;

each R ₁₀ is independently H, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl, or optionally substituted heteroarylalkyl;

each R ₁₁ is independently H, OH, optionally substituted alkyl, optionally substituted alkoxy, or OC(O)R ₁₄ ;

each R ₁₂ is independently H, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl, or optionally substituted heteroarylalkyl;

each R ₁₃ is independently H, OH, optionally substituted alkyl, optionally substituted alkoxy, or OC(O)R ₁₄ ;

each R ₁₄ is independently H, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl, or optionally substituted heteroarylalkyl;

Each t is independently 0, 1, 2, or 3. In another aspect, the salt of the enantiomerically enriched amino alcohol 4b or 4c from step (vi): or a mixture thereof is selected from maleate, malonate, succinate, fumarate, malate, tartrate, dibenzoyl tartrate, di-p-toluoyl tartrate, and mandelate. In another aspect, the salt is tartrate, di-p-toluoyl tartrate, or malate. In another aspect, the salt is L-tartrate, D-di-p-toluoyl tartrate, or D-malate. (Preferably, L-tartrate or D-di-p-toluoyl tartrate).

In another aspect, the present invention provides a method for preparing compound 1 or 1a or a mixture thereof, comprising:

(i) Displace the ester group in ester 2 to obtain morpholine amide 2b:

(ii) Replace the morpholinyl group in morpholinamide 2b: to give ketone 3:

(iii) Formation of the epoxide of ketone 3: to provide epoxide 4:

(iv) Ring opening of epoxide 4: to give amino alcohol ± 4b:

(v) Enantiomeric purity of the enriched amino alcohol ± 4b: to obtain enantiomerically enriched amino alcohol 4b or 4c:

or a mixture thereof;

(vi) arylating the enantiomerically enriched amino alcohol 4b or 4c: or a mixture thereof to obtain the enantiomerically enriched amino alcohol 1-6* or 1-7*: or a mixture thereof;

(vii) forming an enantiomerically enriched salt of amino alcohol 1-6* or 1-7* or a mixture thereof to provide compound XI or Xia: or a mixture thereof;

(viii) generating a tetrazole of XI or Xia: or a mixture thereof to provide compound 1 or 1a: or a mixture thereof;

wherein each R ₁ is independently halogen, —O(C═O)-alkyl, —O(C═O)-substituted alkyl, —O(C═O)-aryl, —O(C═O)-substituted aryl, —O(C═O)-O-alkyl, —O(C═O)-O-substituted alkyl, —O(C═O)-O-aryl, —O(C═O)-O-substituted aryl, —O(SO ₂ )-alkyl, —O(SO ₂ )-substituted alkyl, —O(SO ₂ )-aryl, or —O(SO ₂ )-substituted aryl;

each R ₁₀ is independently H, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl, or optionally substituted heteroarylalkyl;

each R ₁₁ is independently H, OH, optionally substituted alkyl, optionally substituted alkoxy, or OC(O)R ₁₄ ;

each R ₁₂ is independently H, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl, or optionally substituted heteroarylalkyl;

each R ₁₃ is independently H, OH, optionally substituted alkyl, optionally substituted alkoxy, or OC(O)R ₁₄ ;

each R ₁₄ is independently H, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl, or optionally substituted heteroarylalkyl;

Each t is independently 0, 1, 2 or 3. In another aspect, the salt or mixture thereof of the enantiomerically enriched amino alcohol 4b or 4c from step (vii) is selected from maleate, malonate, succinate, fumarate, malate, tartrate, dibenzoyl tartrate, di-p-toluoyl tartrate and mandelate. In another aspect, the salt is tartrate, di-p-toluoyl tartrate or malate. In another aspect, the salt is L-tartrate, D-di-p-toluoyl tartrate or D-malate. (Preferably, L-tartrate or D-di-p-toluoyl tartrate).

In another aspect, Z from any of the embodiments provided herein is phenyl, p-tolyl, methyl, or ethyl.

In another aspect, the crystallization solvent or crystallization solvent mixture from any of the embodiments provided herein is ethyl acetate, isopropyl acetate, ethanol, methanol, or acetonitrile, or a combination thereof.

In another aspect, the crystallization co-solvent or crystallization co-solvent mixture from any of the embodiments provided herein is pentane, methyl tert-butyl ether, hexane, heptane, or toluene, or a combination thereof.

In another aspect, any of the embodiments provided herein can include repeating the enantiomeric enrichment step until the desired level of enantiomeric enrichment is achieved.

In another aspect, Y in any of the embodiments provided herein can be a mesylate or tosylate salt.

In another aspect, any of the embodiments provided herein can include the use of amide 2c in place of morpholine amide 2b.

In another aspect, any of the embodiments provided herein can include replacing ethyl ester 2 with ester 2d.

In other aspects, the invention provides a compound of any of the formulae herein, wherein the compound inhibits (or is believed to inhibit) lanosterol demethylase (CYP51).

In another aspect, the present invention provides a pharmaceutical composition comprising a compound of any of the formulae herein and a pharmaceutically acceptable carrier.

In other aspects, the invention provides a method of modulating metalloenzyme activity in a subject, comprising contacting the subject with a compound of any of the formulae herein in an amount and under conditions sufficient to modulate the metalloenzyme activity.

In one aspect, the present invention provides a method of treating a subject suffering from or susceptible to a condition or disease associated with a metalloenzyme, comprising administering to the subject an effective amount of a compound or pharmaceutical composition of any of the formulae herein.

In another aspect, the present invention provides a method for treating a subject suffering from or susceptible to a disorder or disease associated with a metalloenzyme, wherein the subject has been identified as needing treatment for a disorder or disease associated with a metalloenzyme, the method comprising administering to the subject in need thereof an effective amount of a compound or pharmaceutical composition of any chemical formula herein so that the subject is treated for the disorder.

On the other hand, the present invention provides treatment suffering from or being susceptible to the method for the experimenter of metal enzyme-mediated illness or disease, wherein experimenter has been confirmed as needing the treatment of illness or disease mediated by metal enzyme, the method comprises applying the compound of any chemical formula herein or pharmaceutical composition of effective dose to the experimenter in need, to regulate (for example, lower, suppress) the metal enzyme activity in the experimenter.On the other hand, compound as herein described is more preferably targeted with cancer cell relative to non-transformed cell.

Detailed Description of the Invention

definition

The term "chiral" refers to a molecule that has the property of being non-superimposable with its mirror image, while the term "achiral" refers to a counterpart that is superimposable with its mirror image.

The term "diastereomer" refers to stereoisomers that have two or more asymmetric centers and whose molecules are not mirror images of one another.

The term "enantiomers" refers to two stereoisomers of a compound that are non-superimposable mirror images of one another. An equimolar mixture of two enantiomers is called a "racemic mixture" or a "racemate."

The term "isomers" or "stereoisomers" refers to compounds which have identical chemical constitution, but differ with regard to the arrangement of the atoms or groups in space.

The term "prodrug" includes compounds that have moieties that can be metabolized in vivo. Typically, prodrugs are metabolized in vivo to active drugs by esterases or by other mechanisms. Examples of prodrugs and their uses are well known in the art (see, for example, Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66: 1-19). Prodrugs can be prepared in situ during the final isolation and purification of the compound, or by reacting the purified compound alone in its free acid form or with a hydroxyl group with an appropriate esterifying agent. The hydroxyl group can be converted to an ester by treatment with a carboxylic acid. Examples of prodrug moieties include substituted and unsubstituted, branched and unbranched lower alkyl ester moieties (e.g., propionate), lower alkenyl esters, di-lower alkyl-amino lower alkyl esters (e.g., dimethylaminoethyl ester), amido lower alkyl esters (e.g., acetoxymethyl ester), acyloxy lower alkyl esters (e.g., methyl pivalate), aryl esters (phenyl esters), aryl-lower alkyl esters (e.g., benzyl esters), substituted (e.g., with methyl, halogen, or methoxy substituents) aryl and aryl lower alkyl esters, amides, lower alkyl amides, di-lower alkyl amides, and hydroxyamides. Preferred prodrug moieties are propionate and acyl esters. Prodrugs that are converted to active forms in vivo by other mechanisms are also included. In various aspects, the compounds of the present invention are prodrugs of any of the formulae herein.

The term "subject" refers to an animal, such as a mammal, including but not limited to primates (e.g., humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, etc. In certain embodiments, the subject is a human.

Throughout this application, including the claims, reference to "a sample" without a quantifier refers to "one or more." Thus, for example, reference to "a sample" includes a plurality of samples, etc., unless the context clearly dictates otherwise (e.g., a plurality of samples).

Throughout this specification and claims, unless the context requires otherwise, the word "comprise" is used in its non-exclusive sense.

As used herein, the term "about" when referring to a numerical value is intended to encompass variations of the specified value within, in some embodiments, ±20%, in some embodiments, ±10%, in some embodiments, ±5%, in some embodiments, ±1%, in some embodiments, ±0.5%, and in some embodiments, ±0.1%, as these variations are suitable for practicing the disclosed methods or employing the disclosed compositions.

As used herein, word "inhibitor" means the molecule that shows inhibition metalloenzyme activity. " inhibition " herein means the metalloenzyme activity compared to the absence of inhibitor, and the activity of metalloenzyme reduces. In some embodiments, term " inhibition " means metalloenzyme activity reduction at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or at least about 95%. In other embodiments, inhibition means metalloenzyme activity reduction of about 5% to about 25%, about 25% to about 50%, about 50% to about 70% or about 75% to 100%. In some embodiments, inhibition means metalloenzyme activity reduction of about 95% to 100%, for example, activity reduction of 95%, 96%, 97%, 98%, 99% or 100%. Can use multiple technology well known to those skilled in the art to measure this reduction. The following description is used to measure the specific detection of individual activity.

In addition, the compounds of the present invention include alkenes having any of the following structures: "Z" refers to what is known as the "cis" (same side) configuration, and "E" refers to what is known as the "trans" (opposite side) configuration. With respect to the nomenclature of chiral centers, the terms "d" and "l" configurations are as defined by the IUPAC Recommendations. With respect to the use of the terms diastereomer, racemate, epimer, and enantiomer, these terms are used in their conventional sense to describe the stereochemistry of the formulation.

As used herein, the term "alkyl" refers to a straight or branched hydrocarbon group containing 1 to 12 carbon atoms. The term "lower alkyl" refers to a C1 to C6 alkyl chain. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, tert-butyl, and n-pentyl. Alkyl groups may be optionally substituted with one or more substituents.

The term "alkenyl" refers to an unsaturated hydrocarbon chain, which may be straight or branched, containing from 2 to 12 carbon atoms and at least one carbon-carbon double bond. An alkenyl group may be optionally substituted with one or more substituents.

The term "alkynyl" refers to an unsaturated hydrocarbon chain, which may be straight or branched, containing from 2 to 12 carbon atoms and at least one carbon-carbon triple bond. Alkynyl groups may be optionally substituted with one or more substituents.

The ^sp2 or sp2 carbon of the alkenyl and alkynyl groups can optionally be the point of attachment of the alkenyl and alkynyl groups, respectively.

The term "alkoxy" refers to an -O-alkyl group.

As used herein, the term "halogen," "halo," or "halogenated" refers to -F, -Cl, -Br, or -I.

The term "haloalkoxy" refers to an -O-alkyl group substituted with one or more halo substituents. Examples of haloalkoxy groups include trifluoromethoxy and 2,2,2-trifluoroethoxy.

The term "cycloalkyl" refers to a hydrocarbon having a 3 to 8-membered monocyclic hydrocarbon or a 7 to 14-membered bicyclic ring system with at least one saturated ring or at least one non-aromatic ring, wherein the non-aromatic ring may have a certain degree of unsaturation. The cycloalkyl group may optionally be substituted with one or more substituents. In one embodiment, 0, 1, 2, 3 or 4 atoms of each ring of the cycloalkyl group may be substituted with a substituent. Representative examples of cycloalkyl groups include cyclopropyl, cyclopentyl, cyclohexyl, cyclobutyl, cycloheptyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, etc.

The term "aryl" refers to a monocyclic, bicyclic or tricyclic aromatic hydrocarbon ring system. The aryl group may optionally be substituted with one or more substituents. In one embodiment, 0, 1, 2, 3, 4, 5 or 6 atoms of each ring of the aryl group may be substituted with a substituent. Examples of aryl groups include phenyl, naphthyl, anthracenyl, fluorenyl, indenyl, azulenyl, etc.

The term "heteroaryl" refers to an aromatic 5 to 8 yuan monocycle, an 8 to 12 yuan bicyclic or 11 to 14 yuan tricyclic system, if the system is monocyclic, there is 1 to 4 ring heteroatoms, if the bicyclic, there is 1 to 6 heteroatoms, or if the tricyclic, there is 1 to 9 heteroatoms, the heteroatoms are selected from O, N or S, and the remaining ring atoms are carbon (unless otherwise noted, with suitable hydrogen atoms). Heteroaryl groups can be optionally substituted by one or more substituents. In one embodiment, 0, 1, 2, 3 or 4 atoms in each ring of the heteroaryl group can be substituted by a substituent. The example of heteroaryl groups includes pyridyl, furyl, thienyl, pyrrolyl, oxazolyl, oxadiazolyl, imidazole thiazolyl, isoxazolyl, quinolyl, pyrazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, isoquinolyl, indazolyl etc.

The term "nitrogen-containing heteroaryl" refers to heteroaryl groups having 1 to 4 ring nitrogen heteroatoms if monocyclic, 1 to 6 ring nitrogen heteroatoms if bicyclic, or 1 to 9 ring nitrogen heteroatoms if tricyclic.

The term "heterocycloalkyl" refers to a non-aromatic 3 to 8-membered monocyclic, 7 to 12-membered bicyclic or 10 to 14-membered tricyclic system having 1 to 3 ring heteroatoms if a monocyclic system, 1 to 6 heteroatoms if a bicyclic system, or 1 to 9 heteroatoms if a tricyclic system, wherein the heteroatoms are selected from O, N, S, B, P or Si, and wherein the non-aromatic ring system is fully saturated. The heterocycloalkyl group may optionally be substituted with one or more substituents. In one embodiment, 0, 1, 2, 3 or 4 atoms in each ring of the heterocycloalkyl group may be substituted with a substituent. Representative heterocycloalkyls include piperidinyl, piperazinyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, 1,3-dioxolane, tetrahydrofuranyl, tetrahydrothienyl, thirenyl etc.

The term "alkylamino" refers to an amino substituent further substituted with one or two alkyl groups. The term "aminoalkyl" refers to an alkyl substituent further substituted with one or more amino groups. The term "hydroxyalkyl" or "hydroxyalkyl" refers to an alkyl substituent further substituted with one or more hydroxy groups. The alkyl or aryl portion of alkylamino, aminoalkyl, mercaptoalkyl, hydroxyalkyl, mercaptoalkoxy, sulfonylalkyl, sulfonylaryl, alkylcarbonyl, and alkylcarbonylalkyl groups may be optionally substituted with one or more substituents.

The acids and bases used in the methods herein are known in the art. An acid catalyst is any acidic chemical that can be inorganic (e.g., hydrochloric acid, sulfuric acid, nitric acid, aluminum chloride) or organic (e.g., camphorsulfonic acid, p-toluenesulfonic acid, acetic acid, ytterbium trifluoromethanesulfonate) in nature. An acid is present in a catalytic or stoichiometric amount to facilitate a chemical reaction. A base is any alkaline chemical that can be inorganic (e.g., sodium bicarbonate, potassium hydroxide) or organic (e.g., triethylamine, pyridine) in nature. A base is present in a catalytic or stoichiometric amount to facilitate a chemical reaction.

An alkylating agent is any reagent capable of alkylating the functional group in question (e.g., an oxygen atom in an alcohol, a nitrogen atom in an amino group). Alkylating agents are known in the art, including references cited herein, and include alkyl halides (e.g., methyl iodide, benzyl bromide, or benzyl chloride), alkyl sulfates (e.g., methyl sulfate), or other alkyl leaving group combinations known in the art. Leaving groups are any stable species that can be separated from a molecule during a reaction (e.g., an elimination reaction, a substitution reaction) and are known in the art, including in the references cited herein, and include halides (e.g., I-, Cl-, Br-, F-), hydroxyl groups, alkoxy groups (e.g., -OMe, -Ot-Bu), acyloxy anions (e.g., -OAc, -OC(O) _CF3 ), sulfonates (e.g., mesyl, tosyl), acetamides (e.g., NHC(O)Me), carbamates (e.g., N(Me)C(O)Ot-Bu), phosphates (e.g., -OP(O)(OEt) ₂ )), water, or alcohols (protonated state), and the like.

In certain embodiments, the substituents on any group (such as alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, cycloalkyl, heterocycloalkyl) can be on any atom on the group, wherein any group (such as alkyl, alkenyl, alkynyl, aryl, arylalkyl, heteroaryl, heteroaralkyl, cycloalkyl, heterocycloalkyl) that can be substituted can be optionally substituted by one or more substituents (which can be the same or different) that replace hydrogen atoms respectively. Examples of suitable substituents include but are not limited to alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, halogen, haloalkyl, cyano, nitro, alkoxy, aryloxy, hydroxy, hydroxyalkyl, oxo (i.e., carbonyl), carboxyl, formyl, alkylcarbonyl, alkylcarbonylalkyl, alkoxycarbonyl, alkylcarbonyloxy, aryloxycarbonyl, heteroaryloxy, heteroarylcarbonyloxy, sulfhydryl, sulfhydryl, sulfhydrylalkyl, arylsulfonyl, amino, aminoalkyl, dialkylamino, alkylcarbonylamino , alkylaminocarbonyl, alkoxycarbonylamino, alkylamino, arylamino, diarylamino, alkylcarbonyl, or arylamino substituted aryl; arylalkylamino, aralkylaminocarbonyl, amino, alkylaminosulfonyl, arylaminosulfonyl, dialkylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, imino, urea, carbamoyl, thiourea, thiocyanate, sulfonylamino, sulfonylalkyl, sulfonylaryl, mercaptoalkoxy, N-hydroxyamidino, or N'-aryl, N"-hydroxyamidino.

The compounds of the present invention can be prepared by methods known in the art of organic synthesis. Methods for optimizing reaction conditions and, if necessary, minimizing competing by-products are known in the art. Reaction optimization and scaling up can advantageously employ high-speed parallel synthesis equipment and computer-controlled microreactors (e.g., Design And Optimization in Organic Synthesis, Second Edition, Carlson R, ed., 2005; Elsevier Science Ltd.; K et al., Angew. Chem. Int. Ed. Engl. 2004 43 : 406; and references therein). Other reaction schemes and processes can be determined by the skilled artisan using commercially available searchable structure database software such as (CAS Division of the American Chemical Society) and CrossFire (Elsevier MDL), or by using an internet search engine such as or keyword databases such as the U.S. Patent and Trademark Office text database for appropriate keyword searches. The present invention includes intermediate compounds for preparing the compounds of the chemical formulae herein, as well as methods for preparing these compounds and intermediates, including but not limited to those specifically described in the Examples herein.

The compounds herein may also contain bonds (e.g., carbon-carbon bonds) where rotation of the bond is restricted around that particular bond, such as due to the presence of a ring or double bond. Thus, all cis/trans and E/Z isomers are expressly included in the present invention. The compounds herein may also be represented in multiple tautomeric forms, in which case, although only one tautomeric form may be shown, the present invention expressly includes all tautomeric forms of the compounds described herein. All such isomers of these compounds herein are expressly included in the present invention. All crystalline forms and polymorphs of the compounds described herein are expressly included in the present invention. Also included are extracts and fractions containing the compounds of the present invention. The term isomer is intended to include diastereomers, enantiomers, regioisomers, structural isomers, rotational isomers, tautomers, and the like. For compounds containing one or more stereocenters, such as chiral compounds, the methods of the present invention may be implemented using enantiomerically enriched compounds, racemates, or mixtures of diastereomers.

Preferred enantiomerically enriched compounds have an enantiomeric excess of 50% or more, and more preferably compounds have an enantiomeric excess of 60%, 70%, 80%, 90%, 95%, 98% or 99% or more. In preferred embodiments, only one enantiomer or diastereomer of a chiral compound of the invention is administered to a cell or subject.

Pharmaceutical composition

In one aspect, the present invention provides a pharmaceutical composition comprising a compound of any of the formulae herein and a pharmaceutically acceptable carrier.

In another embodiment, the present invention provides a pharmaceutical composition further comprising other therapeutic agents. In another embodiment, the other therapeutic agent is an anticancer agent, an antifungal agent, a cardiovascular agent, an anti-inflammatory agent, a chemotherapeutic agent, an anti-angiogenic agent, a cytotoxic agent, an antiproliferative agent, a metabolic disease agent, an ophthalmic disease agent, a central nervous system (CNS) disease agent, a urinary system disease agent, or a gastrointestinal disease agent.

In one aspect, the invention provides a test kit comprising an effective amount of a compound of any of the formulas herein in unit dosage form and instructions for administering the compound to a subject suffering from or susceptible to a metalloenzyme-mediated disease or condition, wherein the metalloenzyme-mediated disease or condition comprises cancer, solid tumors, cardiovascular disease, inflammatory disease, infectious disease. In other embodiments, the disease, condition or its symptom is a metabolic disease, an ophthalmic disease, a central nervous system (CNS) disease, a urinary tract disease or a gastrointestinal disease.

The term "pharmaceutically acceptable salt" or "pharmaceutically acceptable carrier" is meant to encompass salts of the active compounds prepared with relatively nontoxic acids or bases, depending on the specific substituents incorporated into the compounds described herein. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of the compound with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salts, or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of the compound with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid or phosphorous acid, and salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, and the like. Also included are salts of amino acids such as arginine, and salts of organic acids such as glucuronic acid or galacturonic acid (see, for example, Berge et al., Journal of Pharmaceutical Science 66: 1-19 (1977)). Certain specific compounds of the present invention contain both basic and acidic functional groups, which enable the compounds to be converted into base addition salts or acid addition salts. Other pharmaceutically acceptable carriers known to those skilled in the art are suitable for use in the present invention.

The neutral form of the compound can be regenerated by contacting the salt with a base or acid and isolating the parent compound using conventional methods. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but the salt and the parent form of the compound are equivalent for the purposes of the present invention.

In addition to salt forms, the present invention provides compounds in the form of prodrugs. Prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide compounds of the present invention. Additionally, prodrugs can be converted into compounds of the present invention by chemical or biochemical methods in vitro. For example, when a prodrug and an appropriate enzyme or chemical reagent are placed in a transdermal patch reservoir, the prodrug can be slowly converted into a compound of the present invention.

Certain compounds of the present invention may exist in unsolvated forms as well as solvated forms, including hydrated forms. Generally speaking, solvated forms are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present invention. Certain compounds of the present invention may exist in polycrystalline or amorphous forms. Generally speaking, all physical forms are equivalent for the uses contemplated by the present invention and are intended to be encompassed within the scope of the present invention.

The present invention also provides pharmaceutical compositions comprising an effective amount of a compound described herein and a pharmaceutically acceptable carrier. In one embodiment, the compound is administered to a subject using a pharmaceutically acceptable dosage form, for example, the pharmaceutically acceptable dosage form can provide sustained release of the compound to the subject for at least 12 hours, 24 hours, 36 hours, 48 hours, one week, two weeks, three weeks, or four weeks after administration of the pharmaceutically acceptable dosage form to the subject.

Actual dosage levels and administration schedules of the active ingredients in the pharmaceutical compositions of the present invention may be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic effect for a particular patient, composition, and mode of administration, without causing toxicity (or unacceptable toxicity) to the patient.

In use, at least one compound according to the present invention is administered in a pharmaceutical carrier in a pharmaceutically effective amount via intravenous, intramuscular, subcutaneous, or intracerebroventricular injection or by oral administration or topical application to a subject in need thereof. According to the present invention, the compound of the present invention can be administered alone or together with another different therapeutic agent. "Together" means together, substantially simultaneously or continuously. In one embodiment, the compound of the present invention is administered acutely. Therefore, the compound of the present invention can be administered in a short course of treatment of, for example, about 1 day to about 1 week. In another embodiment, the compound of the present invention can be administered over a long period of time, for example, about one week to several months depending on the treatment situation to improve chronic conditions.

As used herein, "pharmaceutically effective amount" means an amount of a compound of the invention that is high enough to significantly and positively improve the treatment condition, and low enough to avoid serious side effects (at a reasonable benefit/risk ratio), within the scope of sound medical judgment. A pharmaceutically effective amount of a compound of the invention will vary depending on the specific goals to be achieved, the age and physical condition of the patient to be treated, the severity of the underlying disease, the duration of treatment, the nature of concurrent treatments, and the specific compound used. For example, a therapeutically effective amount of a compound of the invention administered to a child or infant will be proportionally reduced based on sound medical judgment. Thus, an effective amount of a compound of the invention is the minimum amount that will provide the desired effect.

A practical advantage identified with the present invention is that the compounds can be administered in a convenient manner, for example, by intravenous, intramuscular, subcutaneous, oral, or intracerebroventricular injection routes or by topical application, such as a cream or gel. Depending on the route of administration, the active ingredient comprising the compounds of the present invention may need to be coated in a material to protect the compound from enzymes, acids, and other natural conditions that inactivate the compound. In order to administer the compounds of the present invention by non-parenteral administration, the compounds may be coated with or administered with a material to prevent inactivation.

The compounds can be administered parenterally or intraperitoneally. Dispersions can also be prepared in, for example, glycerol, liquid polyethylene glycols, and mixtures thereof and in oils.

Some examples of substances that can be used as pharmaceutical carriers are sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; stearic acid; magnesium stearate; calcium sulfate; vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and cocoa butter; polyols such as propylene glycol, glycerol, sorbitol, mannitol, and polyethylene glycol; agar; alginic acid; pyrogen-free water; isotonic saline; and phosphate buffered saline; skim milk powder; and other non-toxic compatible substances used in pharmaceutical formulations, such as vitamin C, estrogen, and echinacea. Wetting agents and lubricants such as sodium lauryl sulfate may also be present, as well as colorants, flavorings, lubricants, excipients, tableting agents, stabilizers, antioxidants, and preservatives. Solubilizing agents, including, for example, Cremaphore and β-cyclodextrin, may also be used in the pharmaceutical compositions herein.

Pharmaceutical compositions containing the active compound of the presently disclosed subject matter (or its prodrug) can be manufactured by conventional mixing, dissolving, granulating, dragee preparation grinding, emulsifying, encapsulating, encapsulating or lyophilizing processes. The compositions can be prepared by conventional methods using one or more physiologically acceptable carriers, diluents, excipients or adjuvants that facilitate the processing of the active compound into pharmaceutically useful preparations.

The pharmaceutical compositions of the presently disclosed subject matter can be in a form suitable for nearly any route of administration including, for example, topical, ophthalmic, oral, buccal, systemic, nasal, parenteral, transdermal, rectal, vaginal, etc., or in a form suitable for administration by inhalation or insufflation.

For topical administration, the active compound or prodrug can be formulated as solutions, gels, ointments, creams, suspensions, and the like.

Systemic formulations include those designed for administration by injection, for example, subcutaneous, intravenous, intramuscular, intrathecal, or intraperitoneal injection, as well as those designed for transdermal, transmucosal, oral, or pulmonary administration.

Useful injectable formulations include sterile suspensions, solutions or emulsions of the active compound in aqueous or oily media. The composition may also contain a preparaton, such as a suspending agent, a stabilizer and/or a dispersant. The injectable formulation may be presented in unit dose form (e.g., in an ampoule or in a multidose container) and may contain additional preservatives.

Alternatively, the injectable preparation can be provided in powder form for reconstitution with an appropriate medium before use, including but not limited to sterile pyrogen-free water, buffer, glucose solution, etc. To this end, the active compound can be dried by any technique known in the art, such as lyophilization, and reconstituted before use.

For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are known in the art.

For oral administration, the pharmaceutical composition can be in the form of, for example, lozenges, tablets, or capsules, which are prepared by conventional methods using pharmaceutically acceptable excipients such as binders (e.g., pregelatinized corn starch, polyvinyl pyrrolidone, or hydroxypropyl methylcellulose); fillers (e.g., lactose, microcrystalline cellulose, or calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc, or silicon dioxide); dispersants (e.g., potato starch or sodium starch glycolate); or wetting agents (e.g., sodium lauryl sulfate). Tablets can be coated with, for example, sugars or enteric coatings by methods well known in the art.

Liquid preparations for oral administration may be in the form of, for example, elixirs, solutions, syrups, or suspensions, or they may be presented as dry products suitable for reconstitution with water or other appropriate media prior to use. Such liquid preparations may be prepared by conventional methods using pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, cellulose derivatives, or hydrogenated edible fats); emulsifiers (e.g., lecithin or gum arabic); non-aqueous carriers (e.g., almond oil, oily esters, ethanol, or fractionated vegetable oils); preservatives (e.g., methylparaben or propylparaben or sorbic acid). Optionally, the preparation may also contain buffer salts, preservatives, flavorings, coloring agents, and sweeteners.

As is well known, preparations for oral administration may be suitably formulated to provide controlled release of the active compound or prodrug.

For buccal administration, the composition may be in the form of tablets or lozenges formulated in conventional manner.

For rectal and vaginal routes of administration, the active compounds may be formulated as solutions (for retention enemas), suppositories, or ointments containing conventional suppository bases such as cocoa butter or other glycerides.

For nasal administration or administration by inhalation or insufflation, the active compound or prodrug can be conveniently delivered in the form of an aerosol spray from a pressurized pack or nebulizer using a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, fluorocarbons, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to release a metered amount. Capsules and cartridges (e.g., composed of gelatin) for use in an inhaler or insufflator can be formulated as a powder mixture containing the compound and a suitable powder base such as lactose or starch.

A specific example of an aqueous suspension formulation suitable for nasal administration using a commercially available nasal spray device includes the following ingredients: active compound or prodrug (0.5 mg/ml to 20 mg/ml); benzalkonium chloride (0.1 mg/ml to 0.2 mg/mL); polysorbate 80 (80; 0.5 mg/ml to 5 mg/ml); sodium carboxymethylcellulose or microcrystalline cellulose (1 mg/ml to 15 mg/ml); phenylethyl alcohol (1 mg/ml to 4 mg/ml); dextrose (20 mg/ml to 50 mg/ml). The pH of the final suspension can be adjusted to about pH 5 to pH 7, with a typical pH of about pH 5.5.

In order to lastingly deliver, active compound or prodrug can be formulated as the preparation of lasting property of medicine for by subcutaneous injection or intramuscular injection.Active component can be prepared together with suitable polymeric material or hydrophobic material (for example as emulsion in acceptable oil) or ion exchange resin, or be formulated as slightly soluble derivative such as slightly soluble salt.Or, can use the transdermal delivery system of the suction sheet or patch that is manufactured as slow-release active compound for percutaneous absorption.For this reason, penetration enhancer can be used for promoting the transdermal penetration of active compound. Suitable transdermal patches are described, for example, in U.S. Pat. No. 5,407,713, U.S. Pat. No. 5,352,456, U.S. Pat. No. 5,332,213, U.S. Pat. No. 5,336,168, U.S. Pat. No. 5,290,561, U.S. Pat. No. 5,254,346, U.S. Pat. No. 5,164,189, U.S. Pat. No. 5,163,899, U.S. Pat. No. 5,088,977, U.S. Pat. No. 5,087,240, U.S. Pat. No. 5,008,110, U.S. Pat. No. 4,921,475, each of which is incorporated herein by reference in its entirety.

Alternatively, other drug delivery systems may be employed. Liposomes and emulsions are well-known examples of delivery vehicles that can be used to deliver active compounds or prodrugs. Certain organic solvents such as dimethyl sulfoxide (DMSO) may also be employed.

If desired, the pharmaceutical composition may be present in a package or dispenser device that may contain one or more unit dosage forms containing the active compound. For example, the package may comprise metal or plastic film, such as a blister pack. The package or dispenser device may be accompanied by instructions for administration.

The active compounds or prodrugs of the presently disclosed subject matter, or compositions thereof, are generally used in an effective amount to achieve the desired effect, such as an effective amount to treat or prevent the specific disease being treated. The compound can be administered therapeutically to achieve a therapeutic effect, or prophylactically to achieve a preventative effect. A therapeutic effect refers to the eradication or improvement of the underlying disease being treated, and/or the eradication or improvement of one or more symptoms associated with the underlying disease, such that the patient reports an improvement in feeling or health, even though the patient may still be suffering from the underlying disease. For example, administration of a compound to a patient suffering from an allergy provides a therapeutic effect not only when the underlying allergic reaction is eradicated or improved, but also when the patient reports a reduction in the severity or duration of symptoms associated with the allergy following exposure to the allergen. In another example, a therapeutic effect in the case of asthma includes an improvement in breathing after an asthma attack, or a reduction in the severity or frequency of asthma attacks. A therapeutic effect also includes stopping or slowing the progression of the disease, regardless of whether an improvement is achieved.

For preventative administration, this compound can be administered to have and develop into the patient who suffers from one of the aforementioned disease risks.As defined by suitable medical professional or group, having and developing into the patient who suffers from disease risk can be the patient who has the feature that makes the patient place the patient in risk patient's specified group.The patient with risk can also be usually or often in the state that potential disease can develop, and this potential disease can be treated by administration according to metalloenzyme inhibitor of the present invention.In other words, the patient with risk is usually or often exposed under the condition that causes disease or illness, or may expose seriously the patient within a limited time.Perhaps, preventative administration can be used to avoid being diagnosed as the symptom onset of the patient with potential disease.

The amount of compound administered will depend on a variety of factors, including, for example, the specific indication being treated, the mode of administration, whether the desired effect is prophylactic or therapeutic, the severity of the indication being treated, as well as the age and weight of the patient, the bioavailability of the specific compound, etc. The determination of an effective dosage is well within the capabilities of those skilled in the art.

The effective dose can be initially estimated by in vitro testing. For example, the initial dose for animals can be formulated to achieve a circulating blood or serum concentration of the active compound measured in an in vitro test, such as an in vitro fungal MIC or MFC and other in vitro tests described in the Examples section, that is at or above the IC50 of the specific compound. It is within the ability of those skilled in the art to calculate the dose to achieve such circulating blood or serum concentrations, taking into account the bioavailability of the specific compound. For guidance, see Fingl & Woodbury, "General Principles" in The Pharmaceutical Basis of Therapeutics, ed., Goodman & Gilman, Chapter 1, pp. 1-46, latest edition, Pagamonon Press, and the references cited therein, which are incorporated herein by reference.

Initial dosages can also be estimated from in vivo data, such as animal models. Animal models for testing the efficacy of compounds in treating or preventing the various diseases mentioned above are well known in the art.

Dosages generally range from about 0.0001 or 0.001 or 0.01 mg/kg/day to about 100 mg/kg/day, but may also be higher or lower, depending on, among other factors, the activity of the compound, its bioavailability, mode of administration, and the various factors discussed above. Dosages and intervals may be adjusted individually to provide plasma levels of the compound sufficient to maintain a therapeutic or prophylactic effect. In the case of topical administration or selective uptake, such as topical external administration, the effective local concentration of the active compound cannot be correlated with the plasma concentration. Without undue experimentation, the skilled artisan will be able to optimize the effective local dose.

The compounds may be administered once daily, several or more times daily, or even multiple times daily, depending on, among other things, the indication being treated and the judgment of the prescribing physician.

Preferably, the compound will provide a therapeutic or preventive effect without causing substantial toxicity. Standard pharmaceutical procedures can be used to determine the toxicity of a compound. The dose ratio of toxicity to therapeutic effect (or preventive effect) is the therapeutic index. Compounds that exhibit a high therapeutic index are preferred.

The recitation of a list of chemical groups in any definition of a variant herein includes the definition of the variant as any single group or combination of the listed groups. The recitation of an embodiment for a variant herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof. The recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof.

Another object of the present invention is the use of a compound as described herein (e.g., a compound of any formula herein) in the manufacture of a medicament for treating a metalloenzyme-mediated disorder or disease. Another object of the present invention is the use of a compound as described herein (e.g., a compound of any formula herein) in the manufacture of a medicament for treating a metalloenzyme-mediated disorder or disease. Another object of the present invention is the use of a compound as described herein (e.g., a compound of any formula herein) in the manufacture of an agricultural composition for treating or preventing a metalloenzyme-mediated disorder or disease in agriculture or land production.

Agricultural applications

The compounds and compositions herein can be used for methods of regulating metalloenzyme activity in microorganisms on plants, including contacting the compounds (or compositions) herein with plants (e.g., seeds, rice seedlings, grasses, weeds, cereals). By administering the compounds and compositions herein (e.g., contacting, coating, spraying, atomizing, dusting, etc.) to the test plants, land, or other agricultural fields, it can be used to treat plants, land, or other agricultural fields (e.g., as herbicides, pesticides, growth regulators, etc.). The administration can be before or after emergence. The administration can be used as a treatment regimen or a preventive regimen.

Example

The present invention will now be illustrated by using specific examples which are not to be construed as limiting.

General experimental procedures

In the schemes herein, the definitions of the variables in the structures are equivalent to those at corresponding positions in the chemical formulae depicted herein.

Synthesis of 1 or 1a

Disclosed is a method for preparing enantiomeric compounds 1 or 1a. The synthesis of 1 or 1a is achieved using the exemplary synthesis methods shown below (Schemes 1-4). The preparation of the precursor ketone 3-Br begins with the reaction of 2,5-dibromopyridine and ethyl 2-bromodifluoroacetate to prepare the ester 2-Br. The ester can be reacted with morpholine to provide morpholineamide 2b-Br, which is then arylated to provide the ketone 3-Br. Alternatively, as shown in Scheme 1, the ketone 3-Br can be provided directly from the ester 2-Br.

Scheme 1. Synthesis of Ketone 3-Br

In an analogous manner to that described in Scheme 1, ketones 3 (Scheme 2) can be prepared starting from the corresponding substituted 2-bromo-pyridines, which can be prepared according to synthetic transformations known in the art and contained in the references cited herein.

Scheme 2. Synthesis of ketone 3

R ₁ = halogen, -O(C=O)-alkyl, -O(C=O)-substituted alkyl, -O(C=O)-aryl, -O(C=O)-substituted aryl, -O(C=O)-O-alkyl, -O(C=O)-O-substituted alkyl, -O(C=O)-O-aryl, -O(C=O)-O-substituted aryl, -O(SO ₂ )-alkyl, -O(SO ₂ )-substituted alkyl, -O(SO ₂ )-aryl, or -O(SO ₂ )-substituted aryl.

Alternatively, compounds 1 (or 1a, enantiomers of 1, or mixtures thereof) can be prepared using amino alcohols ±4b or ±1-6 according to Scheme 3. Epoxides 4 and 5 can be prepared by reacting ketones 3 and 1-4 with trimethylsulfoxide iodide (TMSI) in the presence of a base (e.g., potassium tert-butoxide) in an appropriate solvent or solvent mixture (e.g., DMSO or THF). Similarly, as shown in Scheme 3, any pyridine compound 3, 4, ±4b, 4b, or 6 can be converted to the corresponding _4- CF3CH2O-Ph analog (e.g., _1-4 , 5, ± _1-6 , _{1-6 *} , or 1, or its corresponding enantiomer, or a mixture thereof) by cross-coupling with 4,4,5,5- _tetramethyl -2-( _4- (2,2,2-tetrafluoroethoxy)phenyl)-1,3,2-dioxaborolane (or the corresponding alkyl borate/ester or _boronic acid, etc.) in the presence of a transition metal catalyst (e.g., (dppf)PdCl2; dppf = 1,1'-(diphenylphosphino _{) ferrocene} ) and a base (e.g., KHCO3, K2CO3, Cs2CO3, or Na2CO3, etc.) in an appropriate solvent system (e.g., an organic aqueous solvent mixture). Epoxides 4 and 5 can then be converted to amino alcohols ±4b and ±1-6 via ammonia-mediated epoxy ring opening using ammonia in a suitable solvent (e.g., MeOH, EtOH, or water). Racemic amino alcohols ±4b and ±1-6 can then be enantiomerically enriched by exposure to chiral acids (e.g., tartaric acid, dibenzoyltartaric acid, or di-p-toluoyltartaric acid, etc.) in a suitable solvent (e.g., acetonitrile, isopropanol, EtOH, or mixtures thereof, or mixtures of any of these with water or MeOH; preferably acetonitrile or a mixture of acetonitrile and MeOH or isopropanol, such as a 90:10, 85:15, or 80:20 mixture). To provide compounds 4b (or 4c, an enantiomer of 4b, or a mixture thereof) or 1-6* (or 1-7*, an enantiomer of 1-6*, or a mixture thereof). Subsequent treatment with TMS-azide in the presence of trimethylorthoformate and sodium acetate in acetic acid affords compound 6 (or 6a, an enantiomer of 6, or a mixture thereof) or 1 (or 1a, an enantiomer of 1, or a mixture thereof) (U.S. Patent No. 4,426,531).

Scheme 3. Synthesis of 1 or 1a via TMSI epoxidation

R ₁ = halogen, -O(C=O)-alkyl, -O(C=O)-substituted alkyl, -O(C=O)-aryl, -O(C=O)-substituted aryl, -O(C=O)-O-alkyl, -O(C=O)-O-substituted alkyl, -O(C=O)-O-aryl, -O(C=O)-O-substituted aryl, -O(SO ₂ )-alkyl, -O(SO ₂ )-substituted alkyl, -O(SO ₂ )-aryl, or -O(SO ₂ )-substituted aryl.

As shown in Scheme 4, compound 1 (or 1a, an enantiomer of 1, or a mixture thereof) prepared by any of the methods disclosed herein can be converted to a sulfonate salt of Formula IX (or IXa, an enantiomer of IX, or a mixture thereof). This can be accomplished by the following steps: a) combining compound 1 (or 1a, an enantiomer of 1, or a mixture thereof), a crystallization solvent or a crystallization solvent mixture (e.g., EtOAc, iPrOAc, EtOH, MeOH, or acetonitrile, or a combination thereof), and a sulfonic acid (e.g., Z═Ph, p-tolyl, Me, or Et), b) diluting the mixture with an appropriate crystallization co-solvent or crystallization co-solvent mixture (e.g., pentane, methyl tert-butyl ether, hexane, heptane, or toluene, or a combination thereof), and c) filtering the mixture to obtain a sulfonate salt of Formula IX (or IXa, an enantiomer of IX, or a mixture thereof).

Scheme 4. Synthesis of Sulfonate Salts of Compounds 1 or 1a

The following describes the HPLC method used to assess the HPLC purity of the Examples and Intermediates shown below:

Column: Waters X Bridge Shield RP 18, 4.6×150mm, 3.5μm

Mobile phase: A = 0.05% TFA/H ₂ O, B = 0.05% TFA/ACN

Autosampler flush: 1:1 ACN/H ₂ O

Diluent: 1:1 ACN/H ₂ O

Flow rate: 1.0 ml/min

Temperature: 45°C

Detector: UV 275nm

Pump parameters:

step Time period A B curve 0 0.5 80.0 20.0 0 1 15.0 60.0 40.0 1 2 10.0 15.0 85.0 1 3 5.0 0.0 100.0 1 4 2.0 0.0 100.0 0 5 8.0 80.0 20.0 0

Example 1

Preparation of ethyl 2-(5-bromopyridin-2-yl)-2,2-difluoroacetate (2-Br)

In a clean, multi-necked round-bottom flask, copper powder (274.7 g, 2.05 eq) was suspended in dimethyl sulfoxide (3.5 L, 7 volumes) at 20° C. to 35° C. Ethyl difluorobromoacetate (449 g, 1.05 eq) was slowly added to the reaction mixture at 20° C. to 25° C. and stirred for 1 to 2 hours. 2,5-Dibromopyridine (500 g, 1 eq) was added to the reaction mixture and the temperature was raised to 35° C. to 40° C. The reaction mixture was maintained at this temperature for 18 to 24 hours and the progress of the reaction was monitored by GC.

After the reaction is complete, ethyl acetate (7 L, 14 volumes) is added to the reaction mixture at 20°C to 35°C and stirring is continued for 60 to 90 minutes. The reaction mixture is filtered through a celite bed (100 g; 0.2 times celite and 1 L; 2 volumes of ethyl acetate). The reactor is washed with ethyl acetate (6 L, 12 volumes) and the washings are filtered through the celite bed. The celite bed is finally washed with ethyl acetate (1 L, 2 volumes) and all filtered mother liquors are combined. The combined ethyl acetate solution is cooled to 8°C to 10°C and washed with buffer solution (5 L, 10 volumes) below 15°C (Note: The addition of buffer solution is naturally exothermic. The buffer solution needs to be added in a controlled manner to maintain the temperature of the reaction mixture below 15°C). The ethyl acetate layer is washed again with buffer solution (7.5 L; 3×5 volumes) until the aqueous layer remains colorless. The organic layer is washed with a 1:1 10 weight/weight % sodium chloride aqueous solution and buffer solution (2.5 L; 5 volumes). The organic layer was then transferred to a dry reactor, and ethyl acetate was distilled under reduced pressure to obtain a crude 2-Br product.

The crude 2-Br product was purified by high vacuum fractional distillation and the distillation fractions containing 2-Br with a purity greater than 93% (with no more than 2% dialkylate and less than 0.5% starting material) were combined to provide 2-Br.

Yield after distillation: 47.7%, purity determined by GC >93% (light yellow liquid).An additional 10% yield was obtained by re-distilling the impure fraction, which brought the total yield to about 55% to 60%.

¹ H NMR: δ values with respect to TMS (DMSO-d ₆ ; 400 MHz): 8.85 (1H, d, 1.6 Hz), 8.34 (1H, dd, J=2.0 Hz, 6.8 Hz), 7.83 (1H, d, J=6.8 Hz), 4.33 (2H, q, J=6.0 Hz), 1.22 (3H, t, J=6.0 Hz). ¹³ CNMR: 162.22 (t, -C=O), 150.40 (Ar-C-), 149.35 (t, Ar-C), 140.52 (Ar-C), 123.01 (Ar-C), 122.07 (Ar-C), 111.80 (t, -CF ₂ ), 63.23 (-OCH ₂ -), 13.45(-CH ₂ CH ₃ ).

Example 2

Preparation of 2-(5-bromopyridin-2-yl)-1-(2,4-difluorophenyl)-2,2-difluoroethanone (3-Br)

A. One-step method

1-Bromo-2,4-difluorobenzene (268.7 g; 1.3 eq) was dissolved in methyl tert-butyl ether (MTBE, 3.78 L, 12.6 vol) at 20°C to 35°C, and the reaction mixture was cooled to -70°C to -65°C using an acetone/dry ice bath. Then, maintaining the reaction temperature below -65°C, n-butyl lithium (689 mL, 1.3 eq; 2.5 M) was added to the reaction mixture (Note: Controlled addition of n-butyl lithium to the reaction mixture was required to maintain the reaction temperature below -65°C). After maintaining the reaction mixture at this temperature for 30 to 45 minutes, 2-Br (300 g, 1 eq) dissolved in MTBE (900 mL, 3 vol) was added to the reaction mixture at below -65°C. The reaction mixture was stirred at this temperature for 60 to 90 minutes, and the progress of the reaction was monitored by GC.

The reaction was quenched by slowly adding 20% w/w ammonium chloride solution (750 mL, 2.5 vol) at below -65°C. The reaction mixture was gradually warmed to 20°C to 35°C, and an additional amount of 20% w/w ammonium chloride solution (750 mL, 2.5 vol) was added. The aqueous layer was separated, and the organic layer was washed with 10% w/w sodium bicarbonate solution (600 mL, 2 vol) and then with 5% sodium chloride (600 mL, 2 vol). The organic layer was dried over sodium sulfate (60 g; 0.2x w/w), filtered, and the sodium sulfate was washed with MTBE (300 mL, 1 vol). The organic layer, along with the washings, was distilled under reduced pressure at below 45°C until no more solvent was collected in the receiving vessel. The distillation temperature was increased to 55-60°C, maintained under vacuum for 3-4 hours, and cooled to 20-35°C to provide 275 g (73.6% yield, 72.71% purity by HPLC) of 3-Br as a light yellow liquid.

¹ H NMR: δ values with respect to TMS (DMSO-d ₆ ; 400 MHz): 8.63 (1H, d, 1.6 Hz, Ar-H), 8.07–8.01 (2H, m, 2×Ar-H), 7.72 (1H, d, J=6.8 Hz, Ar-H), 7.07–6.82 (1H, m, Ar-H), 6.81–6.80 (1H, m, Ar-H). ^13C NMR: 185.60 (t, -C=O), 166.42 (dd, Ar-C-), 162.24 (dd, Ar-C), 150.80 (Ar-C), 150.35 (Ar-C) , 140.02(Ar-C), 133.82(Ar-C), 123.06(Ar-C), 1122.33(Ar-C), 118.44(Ar-C), 114.07(-CF _2- ), 122.07(Ar-C), 105.09(Ar-C).

B. Two-step method via 2b-Br

2-Br (147.0 g) was dissolved in n-heptane (1.21 L) and transferred to a 5 L reactor equipped with an overhead stirrer, thermocouple, condenser, and addition funnel. Morpholine (202 ml) was added. The solution was heated to 60° C. and stirred overnight. HPLC analysis showed that the reaction was complete (0.2% 2-Br; 94.7% 2b-Br). The reaction was cooled to room temperature and 1.21 L of MTBE was added. The solution was cooled to approximately 4° C. and 30% citric acid (563 ml) was slowly added to quench the reaction to maintain an internal temperature of <15° C. After stirring for one hour, the layers were separated (pH of water = 5).

The organic layer was washed with 30% citric acid (322 ml) and 9% _NaHCO₃ (322 ml, pH 7+ of the water after separation). The organic layer was concentrated to 454 g on a rotary evaporator (Note 1) (some precipitate appeared immediately and increased during concentration). After stirring at room temperature, the suspension was filtered and the filter cake was washed with n-heptane (200 ml). The solid was dried in a vacuum oven at room temperature to yield 129.2 g (77%) of a compact powder. The purity determined by HPLC was 96.5%.

To a 1 L flask equipped with an overhead stirrer, condenser, and addition funnel was added magnesium turnings (14.65 g), THF (580 ml), and 1-bromo-2,4-difluorobenzene (30.2 g, 0.39 eq). The mixture was stirred until the reaction began, with spontaneous heating causing the reaction temperature to reach 44°C. The remaining 1-bromo-2,4-difluorobenzene (86.1 g, 1.11 eq) was added over approximately 30 minutes at an internal temperature of 35°C to 40°C, while a cold water bath was used to control the temperature. The reaction was stirred for 2 hours and slowly cooled to room temperature. The dark yellow solution was further cooled to 12°C.

During the Grignard reagent formation, morpholinamide 2b-Br (129.0 g) and THF (645 ml) were charged to a jacketed 2 L flask equipped with an overhead stirrer, a thermocouple, and an addition funnel. The mixture was stirred at room temperature until the solid dissolved, and the solution was then cooled to -8.7°C. The Grignard reagent solution was added via an addition funnel over approximately 30 minutes at a temperature between -5°C and 0°C. The reaction was stirred at 0°C for 1 hour, and the endpoint was determined by HPLC. The reaction mixture was cooled to -5°C and terminated by the addition of 2N HCl at ≤10°C for 1 hour. The mixture was stirred for 0.5 hours, then allowed to separate and the layers were separated. The aqueous layer was extracted with MTBE (280 ml). The combined organic layers were washed with 9% _NaHCO₃ (263 g) and 20% NaCl (258 ml). The organic layer was concentrated on a rotary evaporator and rinsed with THF to transfer all the solution to a distillation flask. Additional THF (100 ml) and toluene (3 x 100 ml) were added and the product was distilled to remove any remaining water. After drying under vacuum, the residue was 159.8 g of a dark brown waxy solid (>theoretical). The purity determined by HPLC was approximately 93%.

Grignard reagent formation/coupling reaction 2:

Magnesium (0.022 kg, 0.903 mole), 1-bromo-2,4-difluorobenzene (0.027 kg, 0.14 mole) and tetrahydrofuran (THF) (1.4 L) were charged into a 2 L reactor equipped with a nitrogen inlet/outlet, a 0.25 L dropping funnel, a temperature probe and a reflux condenser. After stirring at 22° C. for about 40 minutes, the reaction began and was able to reach 35° C. Cooling was implemented, and additional 1-bromo-2,4-difluorobenzene (0.153 kg, 0.79 mole) was added at 35° C. to 40° C. over half an hour. When the addition was complete, the reaction was further stirred at 35° C. to 40° C. for 1 hour, and then the Grignard reagent solution was cooled to 20° C. to 25° C. over 1 hour. During 1 hour cooling, 2b-Br (0.2 kg, 0.62 mole) and THF (0.8 L) were charged into a 5 L reactor equipped with nitrogen inlet/outlet, a 0.5 L dropping funnel, a temperature probe and a reflux condenser, stirred at 15°C to 20°C to obtain a solution, and then cooled to -5°C to 0°C.

At -3 ℃ to 2 ℃, the Grignard reagent was added to the THF solution of morpholineamide over 50 minutes, and the solution was stirred at about 0 ℃ for 1 hour. A sample of the reaction mixture was subjected to GC analysis. A 1 ml sample was quenched in 2M hydrochloric acid solution (5 ml) and extracted with MTBE (2 ml). The organic layer was analyzed, which showed 0.76% morpholineamide residual.

The reaction was terminated by adding 2M hydrochloric acid solution (1 L) at a temperature below 10° C. over 0.75 hours and stirred for a further 0.5 hours. Stirring was terminated and the phases were allowed to separate. The lower aqueous phase was removed and extracted with tert-butyl methyl ether (MTBE) (0.4 L). The combined organic layers were washed with saturated sodium bicarbonate solution (0.4 L) and saturated sodium chloride solution (0.4 L). The solvent was evaporated under vacuum at a temperature below 50° C. and co-distilled with a portion of toluene (0.2 L) until the water content, as determined by Karl Fischer (KF) analysis, was less than 0.1%.

Toluene (0.37 L) and n-heptane (0.37 L) were added to the residue along with SilicaFlash P60 (40 to 63 microns) (0.11 kg) and the reaction was stirred at 20°C to 25°C for 1 hour. The reaction was filtered and washed with toluene/n-heptane (1:1) (2 L). The solvent was evaporated at <50°C and the solvent was switched to THF to provide a 36 wt% 3-Br solution. Gravimetric analysis of a sample of the toluene/n-heptane solution before evaporation showed a mass yield of 0.21 kg (98.5%). GC analysis of the material was 95.34%, giving a contained yield of 93.9%. GC (AUC) analysis of the evaporated sample was 94.5% and HPLC (AUC) analysis gave a purity of 97.1%.

Example 3

Preparation of 5-bromo-2-((2-(2,4-difluorophenyl)oxyethylene-2-yl)difluoromethyl)pyridine (4-Br)

Iodotrimethylsulfoxide (TMSI, 37.93 g; 1.2 eq) was added to a mixture of dimethylsulfoxide (300 mL, 5 vol) and tetrahydrofuran (500 mL, 10 vol) at 20°C to 35°C (a light yellow suspension was observed). Potassium tert-butoxide in THF (172.5 mL, 1.2 eq) was then added to the reaction mixture and stirred at 20°C to 35°C for 60 to 90 minutes to obtain a clear solution. The reaction mixture was then cooled to 0°C to 5°C, maintaining the temperature of the reaction mixture below 15°C, and a solution of 3-Br (50 g, 1 eq) in tetrahydrofuran (150 mL, 3 vol) was added. The progress of the reaction was monitored by GC. The reaction was quenched by the addition of 1 M hydrochloric acid (500 mL, 10 vol) at 0°C to 15°C to bring the pH of the reaction mixture to less than 3. The reaction mixture was maintained at this temperature for 10 to 15 minutes, and then 10% sodium bicarbonate solution (300 mL, 6 volumes) was added to bring the pH of the solution to greater than 7. After maintaining the reaction mixture at 10°C to 15°C for 15 minutes, the reaction mixture was diluted with MTBE (770 mL, 13.5 volumes) and the reaction mixture was allowed to warm to 20°C to 30°C. The organic layer was separated and washed twice with water (100 mL, 2 volumes) and then with 10% sodium chloride (200 mL, 4 volumes). The organic layer was dried over anhydrous sodium sulfate (12.5 g; 0.25 weight/weight), filtered, and the sodium sulfate was washed with MTBE (100 mL, 2 volumes). The filtrate and washings were combined, and the solvent was distilled under reduced pressure below 45°C to provide 35 g (yield 88%, purity determined by GC was >60%) of crude 4-Br product.

The crude 4-Br product was dissolved in MTBE, adsorbed onto silica gel, and purified by silica gel column chromatography using 5% to 10% ethyl acetate in heptane as the mobile phase. The fractions containing 4-Br were combined and the solvent was distilled to provide relatively pure 4-Br. The 4-Br was further purified by slurrying in 4 volumes of 5% ethyl acetate in heptane at room temperature. The pure 4-Br compound was then distilled under reduced pressure below 40°C to provide 15 g (37% yield, >95%) of 4-Br as a light brown solid.

¹ H NMR: δ value for TMS (DMSO-d ₆ ; 400MHz): 8.82 (1H, d, J = 1.6Hz, Ar-H), 8.21 (1H, dd, J = 6.8Hz, 1.6Hz, Ar-H), 7.50 (1H, d, J = 6.8Hz, Ar-H ), 7.43–7.38 (1H, m, Ar-H), 7.27–7.23 (1H, m, Ar-H), 7.11–7.07 (1H, m, Ar-H), 3.39 (1H, d, J = 3.6Hz, -OCH _A H _B -), 3.14 (1H, d, J = 2.0 Hz, -OCH _A H _B -). ¹³ C NMR: 163.87-159.78 (dd, 2×Ar-C-), 150.19 (Ar-C), 149.45 (t, Ar-C), 140.14 (Ar-C), 132.80 (Ar-C), 123.18 (Ar-C), 122.50 (Ar-C), 117.41 (t, -CF ₂ -), 116.71 (Ar-C), 111.58 (Ar-C), 104.04 (t, Ar-C), 57.03 (-CO-CH ₂ -), 49.57 (-CH ₂ -O-).

Second example

Under nitrogen, potassium tert-butoxide (0.061 kg, 0.54 moles; 1.05 equivalents), trimethylsulfoxide iodide (0.125 kg, 0.566 moles; 1.1 equivalents), THF (1.56 L), and dimethylsulfoxide (DMSO) (1.03 L) were added to a 5 L reactor equipped with a nitrogen inlet/outlet, a temperature probe, and a dropping funnel. The mixture was stirred at approximately 20°C for 1 hour (a solution was obtained after approximately 0.25 hours) and then cooled to 0°C. A solution of 3-Br in THF (0.18 kg, 0.53 kg of solution containing 0.515 moles; GC analysis of the solution: 33.6%) was added over 1 hour at 0°C to 2°C and stirred at this temperature for a further 1 hour. A sample of the reaction mixture was taken for GC analysis, which showed a conversion of >99.5% of the starting material.

At 0 ℃ to 5 ℃, by adding 1M hydrochloric acid solution (0.29L) and further continuing stirring for 0.5 hour and quenching reaction.Terminate stirring and make phase separation.The aqueous phase of lower floor is extracted with MTBE (0.58L), and the organic phase of merging is washed with 10% sodium thiosulfate aqueous solution (0.33kg), saturated sodium bicarbonate aqueous solution (0.58L) and saturated sodium chloride solution (0.58L).The weight analysis of solution shows the quantitative mass yield of 4-Br.GC (AUC) analysis is 89.1%, and HPLC (AUC) is 81.5%.Evaporate the solvent under vacuum lower than 50 ℃, replace the solvent with methanol by adding 1.4kg methanol.Further evaporation is implemented under vacuum lower than 50 ℃ to obtain the solid residue (0.189kg) used in the next reaction.

Example 4

Preparation of 3-amino-1-(5-bromopyridin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoropropan-2-ol (±4b-Br)

4-Br (200 g, 1 equivalent) was added to methanolic ammonia (8.0 L; 40 volumes; ammonia content: 15–20% weight/volume) in an autoclave at 10–20°C. Under sealed conditions, the reaction mixture was gradually heated to 60–65°C and 3–4 kg/ ^cm² for 10–12 hours. The progress of the reaction was monitored by GC. After the reaction was complete, the reaction mixture was cooled to 20–30°C and the pressure was gradually released. The solvent was distilled under reduced pressure below 50°C, and the obtained crude product was azeotroped with methanol (2 × 600 mL, 6 volumes) and then isopropanol (600 mL, 2 volumes) to yield 203 g of ±4b-Br (yield 96.98%, purity determined by HPLC: 94.04%).

Second example

4-Br solid (0.18kg, 0.493 mole) and methanol (1.4kg) are fed into a 5L reactor equipped with a nitrogen inlet/outlet, a temperature probe, and a condenser. At 20°C to 25°C, concentrated aqueous ammonia (1.62kg) is added and the reaction is slowly heated to 40°C to 45°C (discharging ammonia). During the heating period, the reaction mixture forms a solution. The solution is further heated for 18 hours, after which a sample is subjected to HPLC analysis, which shows the conversion of the raw material.

The reaction mixture was concentrated to approximately 5 volumes, and MTBE (1.3 L) was added. A 20% aqueous solution of potassium carbonate (0.14 kg) was added (pH 11.5 to 12), and the phases were separated. The upper organic layer was washed with saturated sodium chloride solution (0.29 L). Gravimetric analysis of the MTBE solution (1.09 kg) revealed 0.186 kg of an oil containing ±4b-Br. The purity of the oil was 79.5% by HPLC (AUC) and 82.3% by HPLC (weight/weight) analysis.

Example 5

Preparation of 3-amino-1-(5-bromopyridin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoropropan-2-ol (4b-Br or 2c-Br)

Amino alcohol ± 4b-Br (150 g, 1 equivalent) was dissolved in an isopropanol/acetonitrile mixture (1.5 L, ratio 8:2, 10 volumes) and di-p-toluoyl-L-tartaric acid (L-DPTTA) (84.05 g, 0.55 equivalents) was added to the reactor at 20°C to 30°C. The reaction mixture was heated to 45°C to 50°C for 1 to 1.5 hours (note: the reaction mixture became clear and then became heterogeneous). The reaction mixture was gradually cooled to 20°C to 30°C and stirred for 16 to 18 hours. The resolution process was monitored by chiral HPLC analysis.

After the resolution was complete, the reaction mixture was slowly cooled to 20°C to 35°C. The reaction mixture was filtered, the filter residue washed with a mixture of acetonitrile and isopropanol (8:2 mixture, 300 mL, 2 volumes), and dried to provide 75 g of the L-DPTTA salt (optical purity of 95.37%). The obtained L-DPTTA salt was chirally enriched by suspending the salt in acetonitrile/isopropanol (8:2 mixture, 750 mL, 5 volumes) at 45°C to 50°C for 24 to 48 hours. Chiral enrichment was monitored by chiral HPLC; the solution was gradually cooled to 20°C to 25°C, filtered, and washed with a mixture of isopropanol/acetonitrile (8:2 mixture; 1 volume). This purification process was repeated, and the optical purity of the salt obtained after filtration was greater than 96%. The filtered compound was dried under reduced pressure at 35°C to 40°C to yield 62 g of the enantiomerically enriched L-DPTTA salt as an off-white solid with an optical purity of 97.12%.

The enantiomerically enriched L-DPTTA salt (50 g, 1 equivalent) was dissolved in methanol (150 mL, 3 volumes) at 20°C to 30°C. Potassium carbonate solution (18.05 g of _K₂CO₃ in 150 mL of water) was slowly added with stirring at 20°C to 30° _C . The reaction mixture was maintained at this temperature for 2 to 3 hours (maintaining the pH of the solution at 9). Water (600 mL, 12 volumes) was added to the reaction mixture via an addition funnel, and the reaction mixture was stirred at 20°C to 30°C for 2 to 3 hours. The solid was filtered, rinsed with water (150 mL, 3 volumes), and dried under vacuum at 40°C to 45°C to yield 26.5 g of amino alcohol 4b-Br or 4c-Br as an off-white solid with a chemical purity of 99.54% and an optical purity of 99.28%. (The water content of the chiral amino alcohol was less than 0.10% weight/weight).

¹ H NMR: δ values with respect to TMS (DMSO-d ₆ ; 400 MHz): 8.68 (1H, d, J=2.0 Hz, Ar-H), 8.16 (1H, dd, J=8.0 Hz, 2.0 Hz, Ar-H), 7.49–7.43 (1H, m, Ar-H), 7.40 (1H, d, J=8 Hz, Ar-H), 7.16–7.11 (1H, m, Ar-H), 7.11–6.99 (1H, m, _Ar - _H ), 3.39–3.36 (1H, m, -OCHAHB-), 3.25–3.22 (1H, m, _-OCHAHB- ) _. ^13C NMR: 163.87-158.52(dd, 2×Ar-C-), 150.88(Ar-C), 149.16(Ar-C), 139.21(Ar-C ), 132.39(Ar-C), 124.49(Ar-C), 122.17(Ar-C), 121.87(d, Ar-C), 119.91(t, -CF ₂ -), 110.68 (Ar-C), 103.97 (t, Ar-C), 77.41 (t, -C-OH), 44.17 (-CH ₂ -NH ₂ ).

Second example

Di-p-tolyl-L-tartaric acid (0.069 kg, 0.178 ml; 0.3 eq) was added to a 5 L reactor equipped with a nitrogen inlet/outlet under nitrogen. A solution of ±4b-Br in IPA (1.718 kg; containing 0.225 kg mass of ±4b-Br, 0.59 moles; 1 eq) was added, followed by acetonitrile (0.35 kg). The reaction mixture was stirred at approximately 20°C to obtain a solution. The reaction was heated to 50°C to 55°C (target 52°C) and stirred at this temperature for 4 hours, during which time a precipitate was obtained. A chiral HPLC sample was taken during the reaction by hot filtration of the sample and washed with IPA/acetonitrile (4:1). It showed a chiral purity of >99%.

The reaction was cooled and stirred at 20°C to 25°C for 16 hours. A second sample was subjected to chiral HPLC analysis and was 99.5% pure. The reaction mixture was filtered and washed with a mixture of IPA/acetonitrile (4:1) (0.84 L). The solid obtained was dried under vacuum at 50°C to give 4b-Br hemi-L-DTTA salt as a white solid (0.113 kg). The weight yield was 33.2% and the weight yield of the desired isomer was 66.35%. The purity under chiral HPLC was 99.6% and the purity under achiral HPLC was 99.7%.

Example 6

Preparation of 1-(5-bromopyridin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol (1-6*-Br or 1-7*-Br)

4b-Br or 4c-Br (20.0 g, 1 equivalent) was added to acetic acid (50 mL, 2.5 volumes) at 25°C to 35°C, followed by the addition of anhydrous sodium acetate (4.32 g, 1 equivalent) and trimethyl orthoformate (15.08 g, 2.7 equivalents). The mixture was stirred at this temperature for 15 to 20 minutes, and azidotrimethylsilane (12.74 g, 2.1 equivalents) was added to the reaction mixture (cooling water was circulated through the condenser to reduce the loss of azidotrimethylsilane from the reaction mixture by evaporation). The reaction mixture was then heated to 70°C to 75°C and maintained at this temperature for 2 to 3 hours. The progress of the reaction was monitored by HPLC. Upon completion of the reaction, the reaction mixture was immediately cooled to 25°C to 35°C and water (200 mL, 10 volumes) was added. The reaction mixture was extracted with ethyl acetate (400 mL, 20 volumes), and the aqueous layer was extracted again with ethyl acetate (100 mL, 5 volumes). The combined organic layers were washed with 10% potassium carbonate solution (3 x 200 mL; 3 x 10 volumes) followed by a 10% NaCl wash (1 x 200 mL, 10 volumes). The organic layer was distilled under reduced pressure below 45° C. The crude product obtained was azeotroped with heptane (3 x 200 mL) to afford 21.5 g (94% yield, 99.265% purity) of tetrazole 1-6* or 1-7* compound as a light brown solid (low melting point solid).

¹ H NMR: δ values with respect to TMS (DMSO-d ₆ ; 400 MHz NMR instrument): 9.13 (1H, Ar-H), 8.74 (1H, Ar-H), 8.22–8.20 (1H, m, Ar-H), 7.44 (1H, d, J=7.2 Hz, Ar-H), 7.29 (1H, , Ar-H), 7.23–7.17 (1H, m _, Ar-H), 6.92–6.88 (1H, Ar-H), 5.61 (1H, _d , J=11.2 _Hz , —OCHAHB—), 5.08 (1H, d, J=5.6 Hz, _—OCHAHB— ). ^13C NMR: 163.67-161.59(dd, Ar-C-), 160.60–158.50(dd, Ar-C-), 149.65(Ar-C), 144.99( Ar-C), 139.75 (Ar-C), 131.65 (Ar-C), 124.26 (Ar-C), 122.32 (d, Ar-C), 119.16 (t, -CF ₂ -), 118.70 (d, Ar-C), 111.05 (d, Ar-C), 104.29 (t, Ar-C), 76.79 (t, -C-OH), 59.72 (Ar-C), 50.23 (-OCH ₂ N-).

Example 7

Preparation of 2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-ol (1 or 1a)

A. Preparation of 1 or 1a via 1-6*-Br or 1-7*-Br

Synthesis of 4,4,5,5-tetramethyl-2-(4-(2,2,2-trifluoroethoxy)phenyl)-1,3,2-dioxaborolane

Potassium carbonate (59.7 g, 2.2 eq) was added to a slurry of DMF (190 mL, 3.8 vols), 4-bromophenol (37.4 g, 1.1 eq) and 2,2,2-trifluoroethyl tosylate (50.0 g, 1.0 eq) at 20°C to 35°C under an inert atmosphere. The reaction mixture was heated to 115°C to 120°C and maintained at this temperature for 15 to 18 hours. The progress of the reaction was monitored by GC. The reaction mixture was then cooled to 20°C to 35°C, and toluene (200 mL, 4.0 vols) and water (365 mL, 7.3 vols) were added at the same temperature. The mixture was stirred for 10 to 15 minutes and the layers were separated. The aqueous layer was extracted with toluene (200 mL, 4.0 vols). The organic layers were combined and washed with 2M sodium hydroxide solution (175 mL, 3.5 vols) followed by 20% sodium chloride solution (175 mL, 3.5 vols). The organic layer was then dried over anhydrous sodium sulfate and filtered. The toluene layer was transferred to a clean reactor and purged with argon for at least 1 hour. Bis(pinacolato)diboron (47 g, 1.1 equivalents), potassium acetate (49.6 g, 3.0 equivalents), and 1,4-dioxane (430 mL, 10 volumes) were added at 20°C to 35°C, and the reaction mixture was purged with argon for at least 1 hour. Pd(dppf)Cl2 (6.88 g, 0.05 equivalents) was added to the reaction mixture and the argon purge was continued for 10 to 15 minutes. The temperature of the reaction mixture was raised to 70°C to 75°C and maintained at this temperature under an argon atmosphere for 15 to 35 hours. The reaction progress was monitored by GC. The reaction mixture was cooled to 20°C to 35°C, filtered through a celite pad, and washed with ethyl acetate (86 mL, 2 volumes). The filtrate was washed with water (430 mL, 10 volumes). The aqueous layer was extracted with ethyl acetate (258 mL, 6 volumes), and the combined organic phases were washed with 10% sodium chloride solution (215 mL, 5 volumes). The organic layer was dried over anhydrous sodium sulfate (43 g, 1x weight/weight), filtered, and concentrated under reduced pressure below 45° C. to yield a crude product of 4,4,5,5-tetramethyl-2-(4-(2,2,2-trifluoroethoxy)phenyl)-1,3,2-dioxaborolane (65 g; 71% yield, 85.18% purity as determined by GC). The crude product of 4,4,5,5-tetramethyl-2-(4-(2,2,2-trifluoroethoxy)phenyl)-1,3,2-dioxaborolane (65 g, 1 equivalent) was dissolved in 10% ethyl acetate-n-heptane (455 mL, 7 volumes) and stirred at 20° C. to 35° C. for 30 to 50 minutes. The solution was filtered through a bed of celite and washed with 10% ethyl acetate in n-heptane (195 mL, 3 volumes). The filtrate and washings were combined and concentrated under vacuum at 45°C to afford 4,4,5,5-tetramethyl-2-(4-(2,2,2-trifluoroethoxy)phenyl)-1,3,2-dioxaborolane (45.5 g; 70% recovery) as a viscous slurry. This was then dissolved in 3% ethyl acetate in n-heptane (4 volumes) and adsorbed onto 100M to 200M silica gel (2 times) and eluted through silica gel (4 times) with 3% ethyl acetate in n-heptane. The product-rich fractions were combined and concentrated under vacuum. The column-purified fractions (purity >85%) were transferred to a round-bottom flask equipped with a distillation apparatus. The compound was distilled under high vacuum below 180°C and the fractions were collected. The fractions were analyzed for purity by GC (should have a single largest impurity <1.0% and a purity >98%). Less pure fractions (>85% and <98% purity fractions) were combined and distilled repeatedly to obtain 19 g (32% yield) of light yellow liquid 4,4,5,5-tetramethyl-2-(4-(2,2,2-tetrafluoroethoxy)phenyl)-1,3,2-dioxaborolane.

¹ H NMR: δ values with respect to TMS (DMSO-d ⁶ ; 400 MHz): 7.64 (2H, d, 6.8 Hz), 7.06 (2H, d, J=6.4 Hz), 4.79 (2H, q, J=6.8 Hz), 1.28 (12H, s).

¹³ C NMR: 159.46 (Ar-CO-), 136.24 (2×Ar-C-), 127.77–120.9 (q, -CF ₃ ), 122.0 (Ar-CB), 114.22 (2×Ar-C-), 64.75 (q, J=27.5Hz).

2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy) Synthesis of 2-(phenyl)pyridin-2-yl)propan-2-ol (1 or 1a)

1-6*-Br or 1-7*-Br (14 g, 0.03 mole, 1 eq) was added to tetrahydrofuran (168 mL, 12 vol) at 25°C to 35°C, and the resulting solution was heated to 40°C to 45°C. The reaction mixture was maintained at this temperature for 20 to 30 minutes under argon. Sodium carbonate (8.59 g, 0.08 mole, 2.5 eq) and water (21 mL, 1.5 vol) were added to the reaction mixture, and argon was continued for 20 to 30 minutes. 4,4,5,5-tetramethyl-2-(4-(2,2,2-tetrafluoroethoxy)phenyl)-1,3,2-dioxaborolane (10.76 g, 1.1 eq) dissolved in tetrahydrofuran (42 mL, 3 vol) was added to the reaction mixture, and argon was continued for 20 to 30 minutes. Under argon, Pd(dppf) _Cl₂ (2.65 g, 0.1 equivalent) was added to the reaction mixture and stirred for 20 to 30 minutes (the reaction mixture turned deep red). The reaction mixture was heated to 65 to 70°C and maintained at this temperature for 3 to 4 hours. The progress of the reaction was monitored by HPLC. The reaction mixture was cooled to 40 to 45°C, and the solvent was distilled off under reduced pressure. Toluene (350 mL, 25 vol) was added to the reaction mixture and stirred for 10 to 15 minutes, followed by the addition of water (140 mL, 10 vol). The reaction mixture was filtered through Hyflo (42 g, 3 times), the layers separated, and the organic layer was washed with water (70 mL, 5 vol) and 20% w/w sodium chloride solution (140 mL, 10 vol). The organic layer was treated with charcoal (5.6 g, 0.4 times, neutral charcoal) and filtered through Hyflo. (1S)-10-camphorsulfonic acid (7.2 g, 1 equivalent) was added to the toluene layer and the obtained mixture was heated to 70°C to 75°C for 2 to 3 hours. The reaction mixture was gradually cooled to 25°C to 35°C and stirred for 1 to 2 hours. The solid was filtered, washed with toluene (2×5 volumes), and then dried under vacuum below 45°C to obtain 18.0 g of an off-white solid. The solid (13.5 g, 1 equivalent) was suspended in toluene (135 mL, 10 volumes) and neutralized and stirred for 20 to 30 minutes at 25 to 35°C by adding 1M NaOH solution (1.48 volumes, 1.1 equivalents). Water (67.5 mL, 5 volumes) was added to the reaction mixture and stirred for 10 to 15 minutes, and then the layers were separated. The organic layer was washed with water (67.5 mL, 5 volumes) to remove traces of CSA. Toluene was removed under reduced pressure at 45°C to obtain the crude product of 1 or 1a. After azeotropic removal of traces of toluene with ethanol (3 x 10 volumes), the crude product 1 or 1a was obtained as a light brown solid (7.5 g, 80% yield).

The crude product 1 or 1a (5 g) was dissolved in ethanol (90 mL, 18 volumes) at 20° C. to 35° C. and heated to 40° C. to 45° C. Water (14 volumes) was added to the solution at 40° C. to 45° C., and the solution was maintained at this temperature for 30 to 45 minutes, then gradually cooled to 20° C. to 35° C. The resulting suspension was stirred for an additional 16 to 18 hours at 20° C. to 35° C., and an additional amount of water (4 volumes) was added and stirred for an additional 3 to 4 hours. The solid was filtered to give 4.0 g (80% yield) of 1 or 1a as an off-white solid (HPLC purity >98%).

¹ H NMR: δ value of TMS (DMSO-d ₆ ; 400MHz): 9.15 (1H, s, Ar-H), 8.93 (1H, d, J = 0.8Hz, Ar-H), .8.22–8.20 (1H, m , Ar-H), 7.80 (2H, d, J=6.8Hz, Ar-H), 7.52 (1H, d, J=6.8Hz, Ar-H), 7.29 (1H, d , J＝3.2Hz, Ar-H), 7.27–7.21 (1H, m, Ar-H), 7.23–7.21 (2H, d, J＝6.8Hz, Ar-H) , 7.19 (1H, d, J = 6.8Hz, Ar-H), 6.93–6.89 (1H, m, Ar-H), 5.68 (1H, J = 12Hz, -CH _A H _B ), 5.12 (2H, d, J = 11.6Hz, -CH _A H _B ), 4.85 (2H, q, J = 7.6Hz).

^13C NMR: 163.93–158.33(m, 2xAr-C), 157.56(Ar-C), 149.32(t, Ar-C), 146.40(Ar-C), 145.02(Ar-C), 136. 20(Ar-C), 134.26(2xAr-C), 131.88–131.74(m, AR-C), 129.72(Ar-C), 128.47(2xAr-C), 123.97(q, -CF ₂ -), 122.41(Ar-C), 119.30(-CF ₃ ), 118.99 (Ar-C), 115.65 (2xAr-C), 110.99 (d, Ar-C), 104.22 (t, Ar-C), 77.41–76.80 (m, Ar-C), 64.72 (q, -OCH ₂ -CF ₃ ), 50.54 (-CH ₂ -N-).

B. Preparation of 1 or 1a via 4b-Br or 4c-Br

3-amino-2-(2,4-difluorophenyl)-1,1-difluoro-1-(5-(4-(2,2,2-tetrafluoroethoxy)phenyl)pyridine- Synthesis of 2-amino)propan-2-ol (8a or 8b)

Potassium carbonate (30.4 g) and water (53.3 g) were added to a 1 L flask equipped with overhead stirring, a thermocouple, and a nitrogen/vacuum inlet valve and stirred until dissolved. Boric acid (19.37 g), a 2-butanol solution of 4b-Br or 4c-Br (103.5 g, theoretically 27.8 g of 4b-Br or 4c-Br), and 2-BuOH (147.1 g) were added and stirred to form a clear mixture. The flask was evacuated and refilled with nitrogen three times. Pd(dppf) _2Cl2 (0.30 g) was added and stirred to form _a light orange solution. The flask was evacuated and refilled with nitrogen four times. The mixture was heated to 85°C, stirred overnight, and the endpoint was determined by HPLC analysis. The reaction mixture was cooled to 60°C and the layers separated. The aqueous layer was separated. The organic layer was washed with 5% NaCl solution (5 x 100 ml) at 30°C to 40°C. The organic layer was filtered and transferred to a clean flask rinsed with 2-BuOH. The combined solvent was 309.7 g, with a water content of 13.6 wt % as determined by KF analysis. The solution was diluted with 2-BuOH (189 g) and water (10 g). Theoretically, the solvent contained 34.8 g of product, 522 ml (15 volumes) of 2-BuOH, and 52.2 ml (1.5 volumes) of water. L-tartaric acid (13.25 g) was added, and the mixture was heated to a target temperature of 70°C to 75°C. A thick suspension formed during heating. After maintaining at 70°C to 72°C for about 15 minutes, the suspension became fluid and easy to stir. The suspension was cooled to 25°C at a rate of 10°C/hour and then stirred at 25°C for about 10 hours. The product was collected on a vacuum filter and washed with 10:1 (volume/volume) 2-BuOH/water (50 ml) and 2-butanol (40 ml). The salt was dried in an oven at 60°C under a nitrogen purge for 2 days. 40.08 g of a fluffy, off-white solid 8a or 8b was obtained. The water content was determined by KF analysis to be 0.13 wt %. The yield was 87.3% and the HPLC purity was 99.48%.

2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy) Synthesis of 2-(phenyl)pyridinyl)propan-2-ol (1 or 1a)

Acetic acid (73ml), 8a or 8b (34.8g), sodium acetate (4.58g) and trimethyl orthoformate (16.0g) are added to a 350ml pressure bottle. The mixture is stirred at room temperature for 18 minutes until a uniform suspension is obtained. Azidotrimethylsilane (8.88g) is added and the bottle is sealed. The bottle is immersed in an oil bath and stirred magnetically. The oil bath is initially 52°C and is warmed to 62°C to 64°C in about half an hour. The suspension is stirred overnight at 62°C to 64°C. After 20.5 hours, the suspension is cooled to room temperature and sampled. The reaction is analyzed by HPLC to complete. This reactant and three other reactants using the same raw material pile and general procedure (a total of 3.0g additional raw materials) are combined. The combined reactant is diluted with ethyl acetate (370ml) and water (368ml) and stirred at room temperature for about half an hour. Let stand for stratification, and the layers are separated. The organic layer was washed with 10% _{K2CO3 solution} (370 ml/397 g) and 20% NaCl solution (370 ml/424 g). The organic layer (319 g) was concentrated, diluted with ethanol (202 g) and filtered, rinsing with ethanol (83 g). The combined filtrates were concentrated to 74 g of an amber solution.

The ethanol solution of the crude product of 1 or 1a (74g solution, theoretically containing 31.9g of 1 or 1a ) was transferred to a 2L flask equipped with overhead stirring, a thermocouple and an addition funnel. Ethanol (335g) was added, including ethanol used to complete the solution transfer of 1 or 1a . The solution was heated to a nominal 50°C and water (392g) was added over 12 minutes. The cloudy solution obtained was seeded with 1 or 1a crystals and stirred at 50°C. After about half an hour, the mixture was cooled to 40°C over about half an hour, during which crystallization began. Some dark, short, thick solids separated from the main body of the suspension. The pH of the crystallization mixture was adjusted from 4.5 to 6 using 41% KOH (1.7g). A good suspension was formed after about 1 hour. Additional water (191g) was slowly added over half an hour. The suspension was heated to 50°C and cooled to room temperature at 5°C/minute. After stirring overnight, the suspension was cooled to 16°C in a water bath and filtered after 1 hour. The wet cake was washed with 55:45 (volume/volume) water/ethanol (2×50 ml) and air-dried overnight on a vacuum filter funnel. It was further dried in a vacuum oven at 40° C. with a nitrogen stream to allow no additional weight loss. The product was 30.2 g of a fine off-white powder and some dark particulate matter. The chemical purity of the dark and light materials analyzed by online HPLC was the same. The purity was 99.4%. The water content by KF analysis was 2.16 wt %. The residual ethanol determined by ¹ H NMR analysis was 1.7 wt %. The corrected yield was 29.0 g, with a total yield of 91.0% for tetrazole formation and crystallization. The melting point by DSC analysis was 65° C.

C. Alternative Preparation of 1 or 1a via 4b-Br or 4c-Br

Under nitrogen, 4b-Br semi-L-DTTA salt (0.145 kg, 0.253 mole) and MTBE (0.725 L) were fed to a 5 L reactor equipped with a nitrogen inlet/outlet. The suspension was stirred and a solution of potassium carbonate (0.105 kg, 0.759 mole; 3 equivalents) in water (0.945 kg) was added. The reaction was stirred for 0.25 hours, during which time a solution was obtained. The stirring was stopped and the phases were allowed to separate. The lower aqueous phase (pH 10) was removed and extracted with MTBE (0.725 L). The combined organic layers were evaporated under vacuum at <50°C to give an oil (0.105 kg). 2-Butanol (0.276 kg) was added and distilled to remove residual MTBE. 2-Butanol (0.39 kg) was added. The weight of the 4b-Br(-) solution (0.502 kg) was assumed to theoretically contain free base (0.096 kg) and 2-butanol (0.406 kg).

A solution of potassium carbonate (0.104 kg, 0.759 mole; 3 equivalents) in water (0.184 kg) was prepared and charged to the reactor along with 4-(trifluoroethoxy)phenylboronic acid (0.067 kg, 0.304 mole; 1.2 equivalents). A solution of 4b-Br(-) in 2-butanol was added, followed by further addition of 2-butanol (0.364 kg). The clear solution was purged with nitrogen for 0.5 hour, followed by the addition of Pd(dppf) _Cl catalyst (1.03 g, 0.5 mol %), followed by a continued nitrogen purge for 0.5 hour. The reaction was heated to 85° C. and maintained for 18 hours, after which HPLC IPC analysis showed consumption of the starting material.

The reaction mixture was cooled to 60°C, and the lower aqueous phase was separated (salts precipitated at low temperatures). The organic phase was washed with 5% sodium chloride solution (5 x 0.334 kg) at 30°C to 40°C, during which a small interfacial layer was removed with the final water wash. The organic phase was filtered through a glass fiber filter and washed with 2-butanol (0.065 L). KF analysis of the total solution weight (0.921 kg) gave 15.7% (containing 0.145 kg), assuming theoretically Suzuki free base 7a (0.120 kg) and 2-butanol (0.656 kg). Additional 2-butanol (0.793 kg) and water (0.036 kg) were added. The theoretical reaction composition was 0.120 kg of product, 15 volumes of 2-butanol, and 1.5 volumes of water.

L-Tartaric acid (0.046 kg, 0.304 mol; 1.2 eq) was added and the reaction was heated to 70°C to 75°C. The suspension thickened during heating but thinned out upon reaching temperature. Heating was continued for 1 hour, followed by cooling to 20°C to 25°C at approximately 10°C/hour and stirring for approximately 16 hours. The product was isolated by filtration and washed with 10:1 (v/v) 2-butanol/water (0.17 L) and 2-butanol (0.14 L). The solid was dried under vacuum at 60°C to afford the tartrate ester of 8a as an off-white/grey solid (0.132 kg, 83%). The water content was 2.75% by KF analysis and the purity was 99.5% by HPLC.

Under nitrogen, the tartrate of 8a (0.13 kg, 0.208 mol), sodium acetate (0.017 kg, 0.208 mol), and acetic acid (0.273 L) were charged to a 1 L reactor equipped with a condenser, temperature probe, and nitrogen inlet/outlet. Trimethyl orthoformate (0.132 kg, 1.248 mol; 6 eq) was added, and the suspension was stirred at 20°C to 25°C for 1.25 hours. Azidotrimethylsilane (0.033 kg, 0.287 mol; 1.4 eq) was added to the suspension, and the suspension was heated to 60°C to 65°C and maintained at this temperature for 16 hours. A sample analyzed by HPLC IPC showed 0.2% starting material and 2.9% formamide impurity.

The reaction mixture was cooled to 20°C to 25°C and charged to a 5 L reactor containing ethyl acetate (1.38 L) and purified water (1.38 L). The biphasic solution was stirred for 0.5 hours, and the aqueous phase was removed (pH 4 to 5). A small interfacial layer was retained along with the organic matter. The organic layer was washed with 10% aqueous potassium carbonate (2.2 kg) and separated (water pH 9.3). The organic layer was washed with 20% aqueous sodium chloride (1.625 kg), and the small interfacial layer was removed along with the water.

Under nitrogen conditions, the organic phase was fed to a 2L reactor containing SiliaMetS Thiol palladium detergent (9.2g). The reaction was heated to 50°C to 55°C, maintained at this temperature for 16 hours, and then cooled to 20°C to 25°C. The detergent was removed by filtration through a 0.7 micron filter and washed with ethyl acetate, and the filtrate/washing liquid was evaporated to 100mL under vacuum at <50°C. Ethanol (100%, 755g) was added and the solution was further evaporated to 377g (about 440mL). The solution (theoretical composition is 109g of 1 and 267g of ethanol) was diluted with additional ethanol (1.031kg) and transferred to a 5L reactor. The solution was heated to 50°C and purified water (1.34kg) was added at 45°C to 50°C over 0.25 hours to obtain a turbid solution. It was stirred for 0.5 hours and the pH was adjusted to 6 with 40% potassium carbonate solution (one drop). Stirring was continued at 40°C to 42°C for an additional hour, at which time purified water (0.65 kg) was added over half an hour. The temperature was raised to 50°C and maintained for half an hour, then cooled to 20°C at 10°C/hour. The solid was isolated by filtration and washed with ethanol/water (45:55) (2 x 0.17 L) and dried under vacuum at 45°C to 50°C to yield the X-hydrate of 1 as an off-white solid (0.0937 kg, 85.3%). HPLC (AUC) analysis revealed a purity of 99.62%, containing 0.27% formamide and 0.11% RRT 0.98.

Join by reference

The contents of all references throughout this application (including literature references, issued patents, published patent applications, and co-pending patent applications) are expressly incorporated herein by reference in their entirety.

Equivalent plan

Those skilled in the art will identify or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.

Claims (20)

1. A method for preparing compound 1 or 1a, a mixture thereof, or a salt thereof: It includes morpholinamide 2b: Transformed into compound 1 or 1a, or a mixture thereof; Where _R1 is a halogen, -O(C=O)-alkyl, -O(C=O)-substituted alkyl, -O(C=O)-aryl, -O(C=O)-substituted aryl, -O(C=O)-O-alkyl, -O(C=O)-O-substituted alkyl, -O(C=O)-O-aryl, -O(C=O)-O-substituted aryl, -O( _SO2 )-alkyl, -O( _SO2 )-substituted alkyl, -O( _SO2 )-aryl, or -O( _SO2 )-substituted aryl. 2. The method according to claim 1, comprising making morpholinamide 2b: and The reaction yields compound 1 or 1a or a mixture thereof: Where M is Mg or MgX, Li, _AlX2 ; X is a halogen, alkyl, or aryl group; Where _R1 is a halogen, -O(C=O)-alkyl, -O(C=O)-substituted alkyl, -O(C=O)-aryl, -O(C=O)-substituted aryl, -O(C=O)-O-alkyl, -O(C=O)-O-substituted alkyl, -O(C=O)-O-aryl, -O(C=O)-O-substituted aryl, -O( _SO2 )-alkyl, -O( _SO2 )-substituted alkyl, -O( _SO2 )-aryl, or -O( _SO2 )-substituted aryl. 3. The method according to claim 2, wherein M is Mg or MgX, and X is a halogen. 4. The method according to claim 1, further comprising amidation of ester 2: To obtain morpholinamide 2b: Each _R1 is independently a halogen, -O(C=O)-alkyl, -O(C=O)-substituted alkyl, -O(C=O)-aryl, -O(C=O)-substituted aryl, -O(C=O)-O-alkyl, -O(C=O)-O-substituted alkyl, -O(C=O)-O-aryl, -O(C=O)-O-substituted aryl, -O( _SO₂ )-alkyl, -O( _SO₂ )-substituted alkyl, -O( _SO₂ )-aryl, or -O( _SO₂ )-substituted aryl. 5. The method according to claim 4, further comprising reacting ester 2 with morpholine: To obtain morpholinamide 2b: Each _R1 is independently a halogen, -O(C=O)-alkyl, -O(C=O)-substituted alkyl, -O(C=O)-aryl, -O(C=O)-substituted aryl, -O(C=O)-O-alkyl, -O(C=O)-O-substituted alkyl, -O(C=O)-O-aryl, -O(C=O)-O-substituted aryl, -O( _SO₂ )-alkyl, -O( _SO₂ )-substituted alkyl, -O( _SO₂ )-aryl, or -O( _SO₂ )-substituted aryl. 6. The method according to claim 1, further comprising: (i) Replace the morpholino group in morpholinamide 2b to obtain ketone 3: (ii) Aryl ketone 3: to obtain aryl-pyridine 1-4: (iii) An epoxide of aryl-pyridine 1-4 is formed to obtain epoxide 5. (iv) Ring-opening of epoxide 5: to obtain amino alcohol ±1-6: (v) Enrich the enantiomeric purity of amino alcohols ±1-6: to obtain enantiomeric enriched amino alcohols 1-6* or 1-7*: or mixtures thereof; (vi) Generate enantiomeric enriched amino alcohols 1-6* or 1-7*: or mixtures thereof in tetrazolium to obtain compound 1 or 1a: or mixtures thereof; Each _R1 is independently a halogen, -O(C=O)-alkyl, -O(C=O)-substituted alkyl, -O(C=O)-aryl, -O(C=O)-substituted aryl, -O(C=O)-O-alkyl, -O(C=O)-O-substituted alkyl, -O(C=O)-O-aryl, -O(C=O)-O-substituted aryl, -O( _SO₂ )-alkyl, -O( _SO₂ )-substituted alkyl, -O( _SO₂ )-aryl, or -O( _SO₂ )-substituted aryl. 7. The method according to claim 1, further comprising: (i) Substitution of the morpholino group in morpholinamide 2b yields ketone 3: (ii) An epoxide of ketone 3 is formed, yielding epoxide 4. (iii) Ring-opening of epoxide 4: yields amino alcohol ±4b: (iv) Enantiomeric purity of amino alcohol ± 4b: to obtain enantiomeric enriched amino alcohol 4b or 4c: or mixtures thereof; (v) Generate enantiomeric enriched amino alcohols 4b or 4c: Tetraazoles of or mixtures thereof yield tetraazole 6 or 6a: or mixtures thereof; (vi) Aryltetrazole 6 or 6a: or a mixture thereof, to obtain compound 1 or 1a: or a mixture thereof; Each _R1 is independently a halogen, -O(C=O)-alkyl, -O(C=O)-substituted alkyl, -O(C=O)-aryl, -O(C=O)-substituted aryl, -O(C=O)-O-alkyl, -O(C=O)-O-substituted alkyl, -O(C=O)-O-aryl, -O(C=O)-O-substituted aryl, -O( _SO₂ )-alkyl, -O( _SO₂ )-substituted alkyl, -O( _SO₂ )-aryl, or -O( _SO₂ )-substituted aryl. 8. The method according to claim 6 or 7, further comprising: (i) Combining compound 1 or 1a: or a mixture thereof, sulfonic acid and a crystallizing solvent or a mixture of crystallizing solvents; (ii) Dilute the mixture from step (i) using a crystallization cosolvent or a mixture of crystallization cosolvents; and (iii) Compounds of the isolated form IX or IXa: or mixtures thereof; Each Z is independently an aryl, substituted aryl, alkyl, or substituted alkyl. 9. The method according to claim 8, wherein Z is phenyl, p-tolyl, methyl, or ethyl. 10. The method of claim 8, wherein the crystallization solvent or mixture of crystallization solvents is ethyl acetate, isopropyl acetate, ethanol, methanol, or acetonitrile, or a combination thereof. 11. The method according to claim 8, wherein the crystallization cosolvent or mixture of crystallization cosolvents is pentane, methyl tert-butyl ether, hexane, heptane, or toluene or a combination thereof. 12. The method according to claim 1, further comprising: (i) Replacing the ester group of ester 2: yields morpholinamide 2b: (ii) Substitution of the morpholino group in morpholinamide 2b yields ketone 3: (iii) An epoxide of ketone 3 is formed, yielding epoxide 4. (iv) Ring-opening of epoxide 4: yields amino alcohol ± 4b: (v) Enantiomeric purity of enriched amino alcohols ± 4b: to obtain enantiomeric enriched amino alcohols 4b or 4c: or mixtures thereof; (vi) Arylation of enantiomerically enriched amino alcohols 4b or 4c: or mixtures thereof, to obtain enantiomerically enriched amino alcohols 1-6* or 1-7*: or mixtures thereof; (vii) Salts of enantiomeric enriched amino alcohols 1-6* or 1-7*, or mixtures thereof, are generated to give compounds XI or Xia, or mixtures thereof; (viii) Generate a tetrazolium of XI or Xia: or a mixture thereof to obtain compound 1 or 1a: or a mixture thereof; Each _R1 is independently a halogen, -O(C=O)-alkyl, -O(C=O)-substituted alkyl, -O(C=O)-aryl, -O(C=O)-substituted aryl, -O(C=O)-O-alkyl, -O(C=O)-O-substituted alkyl, -O(C=O)-O-aryl, -O(C=O)-O-substituted aryl, -O( _SO₂ )-alkyl, -O( _SO₂ )-substituted alkyl, -O( _SO₂ )-aryl, or -O( _SO₂ )-substituted aryl. 13. The method of claim 12, wherein the salt is selected from maleate, malonate, succinate, fumarate, malate, tartrate, dibenzoyl tartrate, ditolyl tartrate, and mandelate. 14. The method of claim 12, wherein the salt is a tartrate, di-toluyl tartrate, or malate. 15. The method of claim 12, wherein the salt is L-tartrate or D-malate. 16. The method of claim 12, wherein the salt is an L-tartrate. 17. A method for preparing epoxide 5:, the method comprising: (i) Replace the morpholino group in morpholinamide 2b to obtain ketone 3: (ii) Aryl ketone 3: to obtain aryl-pyridine 1-4: (iii) To generate 1-4 epoxides of aryl-pyridine to obtain epoxide 5: Each _R1 is independently a halogen, -O(C=O)-alkyl, -O(C=O)-substituted alkyl, -O(C=O)-aryl, -O(C=O)-substituted aryl, -O(C=O)-O-alkyl, -O(C=O)-O-substituted alkyl, -O(C=O)-O-aryl, -O(C=O)-O-substituted aryl, -O( _SO₂ )-alkyl, -O( _SO₂ )-substituted alkyl, -O( _SO₂ )-aryl, or -O( _SO₂ )-substituted aryl. 18. A method for preparing enantiomerically enriched amino alcohols 1-6* or 1-7*, or mixtures thereof, the method comprising: (i) Replace the morpholino group in morpholinamide 2b to obtain ketone 3: (ii) Aryl ketone 3: to obtain aryl-pyridine 1-4: (iii) To generate 1-4 epoxides of aryl-pyridine to obtain epoxide 5: (iv) Ring-opening of epoxide 5: to obtain amino alcohol ±1-6: (v) Enrich the enantiomeric purity of amino alcohols ±1-6: to obtain enantiomeric enriched amino alcohols 1-6* or 1-7*: or mixtures thereof; Each _R1 is independently a halogen, -O(C=O)-alkyl, -O(C=O)-substituted alkyl, -O(C=O)-aryl, -O(C=O)-substituted aryl, -O(C=O)-O-alkyl, -O(C=O)-O-substituted alkyl, -O(C=O)-O-aryl, -O(C=O)-O-substituted aryl, -O( _SO₂ )-alkyl, -O( _SO₂ )-substituted alkyl, -O( _SO₂ )-aryl, or -O( _SO₂ )-substituted aryl. 19. A method for preparing enantiomerically enriched amino alcohols 1-6* or 1-7*, or mixtures thereof, the method comprising: (i) Substitution of the morpholino group in morpholinamide 2b yields ketone 3: (ii) An epoxide of ketone 3 is formed, yielding epoxide 4. (iii) Ring-opening of epoxide 4: yields amino alcohol ±4b: (iv) Enantiomeric purity of amino alcohol ± 4b: to obtain enantiomeric enriched amino alcohol 4b or 4c: or mixtures thereof; (v) Arylation of enantiomeric enriched amino alcohols 4b or 4c: or mixtures thereof, to obtain enantiomeric enriched arylpyridines 1-6* or 1-7*: or mixtures thereof; Each _R1 is independently a halogen, -O(C=O)-alkyl, -O(C=O)-substituted alkyl, -O(C=O)-aryl, -O(C=O)-substituted aryl, -O(C=O)-O-alkyl, -O(C=O)-O-substituted alkyl, -O(C=O)-O-aryl, -O(C=O)-O-substituted aryl, -O( _SO₂ )-alkyl, -O( _SO₂ )-substituted alkyl, -O( _SO₂ )-aryl, or -O( _SO₂ )-substituted aryl. 20. A method for preparing enantiomerically enriched amino alcohols 4b or 4c: A method for a mixture thereof, the method comprising: (i) Substitution of the morpholino group in morpholinamide 2b yields ketone 3: (ii) To generate the epoxide of ketone 3: to obtain epoxide 4: (iii) Ring-opening of epoxide 4: yields amino alcohol ±4b: (iv) Enantiomeric purity of amino alcohol ± 4b: to obtain enantiomeric enriched amino alcohol 4b or 4c: or mixtures thereof; Each _R1 is independently a halogen, -O(C=O)-alkyl, -O(C=O)-substituted alkyl, -O(C=O)-aryl, -O(C=O)-substituted aryl, -O(C=O)-O-alkyl, -O(C=O)-O-substituted alkyl, -O(C=O)-O-aryl, -O(C=O)-O-substituted aryl, -O( _SO₂ )-alkyl, -O( _SO₂ )-substituted alkyl, -O( _SO₂ )-aryl, or -O( _SO₂ )-substituted aryl.