U.S. patent application number 16/916108 was filed with the patent office on 2020-10-22 for sglts inhibitor and application thereof.
This patent application is currently assigned to SHANDONG DANHONG PHARMACEUTICAL CO., LTD.. The applicant listed for this patent is SHANDONG DANHONG PHARMACEUTICAL CO., LTD.. Invention is credited to SHUHUI CHEN, YI LI, QINGHUA MAO, CHENGDE WU, TAO YU.
Application Number | 20200331950 16/916108 |
Document ID | / |
Family ID | 1000004969125 |
Filed Date | 2020-10-22 |
![](/patent/app/20200331950/US20200331950A1-20201022-C00001.png)
![](/patent/app/20200331950/US20200331950A1-20201022-C00002.png)
![](/patent/app/20200331950/US20200331950A1-20201022-C00003.png)
![](/patent/app/20200331950/US20200331950A1-20201022-C00004.png)
![](/patent/app/20200331950/US20200331950A1-20201022-C00005.png)
![](/patent/app/20200331950/US20200331950A1-20201022-C00006.png)
![](/patent/app/20200331950/US20200331950A1-20201022-C00007.png)
![](/patent/app/20200331950/US20200331950A1-20201022-C00008.png)
![](/patent/app/20200331950/US20200331950A1-20201022-C00009.png)
![](/patent/app/20200331950/US20200331950A1-20201022-C00010.png)
![](/patent/app/20200331950/US20200331950A1-20201022-C00011.png)
View All Diagrams
United States Patent
Application |
20200331950 |
Kind Code |
A1 |
WU; CHENGDE ; et
al. |
October 22, 2020 |
SGLTS INHIBITOR AND APPLICATION THEREOF
Abstract
A compound as an SGLT1/SGLT2 dual inhibitor, and an application
thereof in the preparation of a drug as the SGLT1/SGLT2 dual
inhibitor. The compound is a compound represented by formula (I),
an isomer thereof, or a pharmaceutically acceptable salt thereof.
##STR00001##
Inventors: |
WU; CHENGDE; (SHANGHAI,
CN) ; MAO; QINGHUA; (SHANGHAI, CN) ; LI;
YI; (SHANGHAI, CN) ; YU; TAO; (SHANGHAI,
CN) ; CHEN; SHUHUI; (SHANGHAI, CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
SHANDONG DANHONG PHARMACEUTICAL CO., LTD. |
Heze |
|
CN |
|
|
Assignee: |
SHANDONG DANHONG PHARMACEUTICAL
CO., LTD.
|
Family ID: |
1000004969125 |
Appl. No.: |
16/916108 |
Filed: |
June 30, 2020 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
PCT/CN2019/070336 |
Jan 4, 2019 |
|
|
|
16916108 |
|
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07H 15/14 20130101;
A61P 3/10 20180101 |
International
Class: |
C07H 15/14 20060101
C07H015/14; A61P 3/10 20060101 A61P003/10 |
Foreign Application Data
Date |
Code |
Application Number |
Jan 5, 2018 |
CN |
201810012284.4 |
Claims
1. A compound represented by formula (I), an isomer thereof, or a
pharmaceutically acceptable salt thereof, ##STR00067## wherein, m
is 1 or 2; n is 0, 1 or 2; D is --O-- or --C(R.sub.1)(R.sub.2)--;
ring A is selected from phenyl and 5-6 membered heteroaryl; R.sub.1
is selected from the group consisting of H, F, Cl, Br, I, OH and
NH.sub.2; R.sub.2 is selected from the group consisting of H, F,
Cl, Br and I; or R.sub.1 and R.sub.2 are connected to form a 5-6
membered heterocycloalkyl; R.sub.3 is selected from the group
consisting of H, F, Cl, Br, I, OH, NH.sub.2, C.sub.1-3 alkyl and
C.sub.1-3 alkoxy, wherein the C.sub.1-3 alkyl and C.sub.1-3 alkoxy
are optionally substituted by one, two or three R group(s); R.sub.4
is selected from C.sub.1-3 alkyl, wherein the C.sub.1-3 alkyl is
optionally substituted by one, two or three R group(s); R is
selected from the group consisting of F, Cl, Br, I, OH and
NH.sub.2; and the 5-6 membered heteroaryl and 5-6 membered
heterocycloalkyl respectively contain one, two, three or four
heteroatom(s) or heteroatom group(s) independently selected from
the group consisting of --NH--, --O--, --S-- and N.
2. The compound, the isomer thereof, or the pharmaceutically
acceptable salt thereof according to claim 1, wherein R.sub.3 is
selected from the group consisting of H, F, Cl, Br, I, OH,
NH.sub.2, CH.sub.3, Et, and --O--CH.sub.3.
3. The compound, the isomer thereof, or the pharmaceutically
acceptable salt thereof according to claim 1, wherein R.sub.4 is
selected from CH.sub.3 and Et.
4. The compound, the isomer thereof, or the pharmaceutically
acceptable salt thereof according to claim 1, wherein the ring A is
selected from phenyl and thienyl.
5. The compound, the isomer thereof, or the pharmaceutically
acceptable salt thereof according to claim 4, wherein the ring A is
selected from ##STR00068##
6. The compound, the isomer thereof, or the pharmaceutically
acceptable salt thereof according to claim 1, wherein the
structural unit ##STR00069## is selected from ##STR00070##
7. The compound, the isomer thereof, or the pharmaceutically
acceptable salt thereof according to claim 6, wherein the
structural unit ##STR00071## is selected from ##STR00072##
8. The compound, the isomer thereof, or the pharmaceutically
acceptable salt thereof according to claim 1, wherein the
structural unit ##STR00073## is selected from ##STR00074##
9. The compound, the isomer thereof, or the pharmaceutically
acceptable salt thereof according to claim 1, selected from the
group consisting of ##STR00075## wherein, R.sub.1 and R.sub.2 are
as defined in claim 1; R.sub.3 is as defined in claim 1; R.sub.4 is
as defined in claim 1.
10. The compound of the following formula, an isomer thereof, or a
pharmaceutically acceptable salt thereof according to claim 1,
selected from ##STR00076## ##STR00077##
11. A pharmaceutical composition, comprising a therapeutically
effective amount of a compound, an isomer thereof or a
pharmaceutically acceptable salt thereof according to claim 1 as an
active ingredient, and a pharmaceutically acceptable carrier.
12. A method for treating SGLT1/SGLT2 related diseases, comprising
a step of administering the compound or a pharmaceutically
acceptable salt thereof according to claim 1 to a subject in
need.
13. The method according to claim 12, wherein the disease is
diabetes.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of International Patent
Application No. PCT/CN2019/070336 with a filing date of Jan. 4,
2019, designating the United States, now pending, and claims
priority to Chinese Patent Application No. 201810012284.4 with a
filing date of Jan. 5, 2018. The content of the aforementioned
applications, including any intervening amendments thereto, are
incorporated herein by reference.
TECHNICAL FIELD
[0002] The present invention relates to a compound as an
SGLT1/SGLT2 dual inhibitor, and an application thereof in the
preparation of a drug as the SGLT1/SGLT2 dual inhibitor.
Specifically, the compound is a compound represented by formula
(I), an isomer thereof, or a pharmaceutically acceptable salt
thereof.
BACKGROUND
[0003] Diabetes is a metabolic disease characterized by
hyperglycemia. Hyperglycemia is caused by deficiency of insulin
secretion and/or damage of biological function. In patients with
diabetes, long-term abnormal blood glucose levels can lead to
serious complications, including cardiovascular disease, chronic
renal failure, retinal injury, nerve injury, microvascular injury
and obesity or like. In the early stage of the treatment of
diabetes, diet control and exercise therapy are the preferred
methods for blood glucose control. When these methods do not work,
insulin or oral hypoglycemic drugs are required for the treatment.
At present, various hypoglycemic drugs, including biguanides,
sulfonylureas, insulin resistance improvers, glinides,
.alpha.-glucosidase inhibitors, dipeptidyl peptidase-IV inhibitors
and the like, are used in clinical treatment. These drugs have good
therapeutic effects, but also have safety problems in the long-term
treatment. For example, biguanides may cause lactic acidosis,
sulfonylureas may cause hypoglycemia, insulin resistance improvers
may cause edema, heart failure and weight gain, and
.alpha.-glucosidase inhibitors may cause symptoms including
abdominal pain, abdominal distension, diarrhea or like. Therefore,
there is an urgent need to develop a safer and more effective new
hypoglycemic drug to meet the needs of the treatment of
diabetes.
[0004] Sodium-glucose cotransporters (SGLTs) are a family of
glucose transporters found in the small intestinal mucosa and renal
proximal convoluted tubules. The family mainly comprises two
members including SGLT1 proteins and SGLT2 proteins which mediate
the transmembrane transport of glucose in the intestine and kidney
and play a key role in maintaining the stability of human blood
glucose. Specifically, SGLT1 is distributed mainly in intestinal
mucosal cells of the small intestine and is also expressed in a
small amount in myocardium and kidney, and mainly regulates the
absorption of glucose in the intestine. SGLT2 is expressed at a
high level in the kidney, mainly responsible for the regulation of
glucose reuptake in the kidney, that is, when glucose in urine is
filtered through glomerulus, it can be actively attached to renal
tubular epithelial cells and transported into cells through SGLT2
protein so that the glucose can be reused. In this process, SGLT2
is responsible for the reabsorption of 90% glucose, and the
reabsorption of the remaining 10% is done by SGLT1. This process is
not involved in glucose metabolism, thus avoiding or reducing the
occurrence of adverse reactions of hypoglycemia, and further
reducing the risk of cardiovascular diseases. Therefore, SGLTs has
become one of the ideal potential targets for the treatment of
diabetes.
[0005] In view of this, some SGLTs inhibitors, especially highly
selective SGLT2 inhibitors, have been developed successively. These
inhibitors inhibit the activity of SGLT2, and thereby specifically
inhibit the reabsorption of glucose by the kidney, thus increasing
the excretion of glucose in urine and normalizing plasma glucose in
patients with diabetes. Since 2012, six drugs including
Dapagliflozin, Canagliflozin, Luseogliflozin, Ipragliflozin,
Tofogliflozin and Empagliflozin have been approved for marketing
and have become effective drugs for the treatment of diabetes.
[0006] In addition to use of selective SGLT2 inhibitors, recent
studies have shown that inhibition of SGLT2 and partial inhibition
of SGLT1 can not only inhibit glucose reuptake in the kidney, but
also control the absorption of glucose in the intestine without
diarrhea or other gastrointestinal reactions. Furthermore, it is
found that inhibition of SGLT1 in the intestine can reduce the
amount of glucose entering the blood from the gastrointestinal
tract, increase levels of GLP-1 and PYY after meals, show a better
hypoglycemic effect than selective SGLT2 inhibitors and can reduce
the risk of urinary tract infection and renal function damage.
Therefore, the development of SGLT1/SGLT2 dual inhibitor has become
a new target and direction for the treatment of diabetes in recent
years.
[0007] In summary, as a new type drug for the treatment of
diabetes, the SGLT1/SGLT2 dual inhibitor has a good development
prospect. Therefore, there is an urgent need to develop a
SGLT1/SGLT2 dual inhibitor with excellent efficacy, good
pharmacokinetics and high safety for the treatment of diabetes and
related metabolic disorders. At present, Lexicon and Sanofi have
jointly developed a SGLT1/SGLT2 dual inhibitor named Sotagliflozin
and the Phase III clinical study has been completed
(WO2008042688/WO2012094293).
##STR00002##
SUMMARY
[0008] The present disclosure provides a compound represented by
formula (I), an isomer thereof, or a pharmaceutically acceptable
salt thereof,
##STR00003##
[0009] wherein, m is 1 or 2;
[0010] n is 0, 1 or 2;
[0011] D is --O-- or --C(R.sub.1)(R.sub.2)--;
[0012] ring A is selected from phenyl and 5-6 membered
heteroaryl;
[0013] R.sub.1 is selected from the group consisting of H, F, Cl,
Br, I, OH and NH.sub.2;
[0014] R.sub.2 is selected from the group consisting of H, F, Cl,
Br and I;
[0015] or R.sub.1 and R.sub.2 are connected to form a 5-6 membered
heterocycloalkyl;
[0016] R.sub.3 is selected from the group consisting of H, F, Cl,
Br, I, OH, NH.sub.2, C.sub.1-3 alkyl and C.sub.1-3 alkoxy, wherein
the C.sub.1-3 alkyl and C.sub.1-3 alkoxy are optionally substituted
by one, two or three R group(s);
[0017] R.sub.4 is selected from C.sub.1-3 alkyl, wherein the
C.sub.1-3 alkyl is optionally substituted by one, two or three R
group(s);
[0018] R is selected from the group consisting of F, Cl, Br, I, OH
and NH.sub.2; and
[0019] the 5-6 membered heteroaryl and 5-6 membered
heterocycloalkyl respectively contain one, two, three or four
heteroatom(s) or heteroatom group(s) independently selected from
the group consisting of --NH--, --O--, --S-- and N.
[0020] In some embodiments of the present disclosure, the above
R.sub.3 is selected from the group consisting of H, F, Cl, Br, I,
OH, NH.sub.2, CH.sub.3, Et, and --O--CH.sub.3. Other variables are
as defined by the present disclosure.
[0021] In some embodiments of the present disclosure, the above
R.sub.4 is selected from CH.sub.3 and Et. Other variables are as
defined by the present disclosure.
[0022] In some embodiments of the present disclosure, the above
ring A is selected from phenyl and thienyl. Other variables are as
defined by the present disclosure.
[0023] In some embodiments of the present disclosure, the above
ring A is selected from
##STR00004##
Other variables are as defined by the present disclosure.
[0024] In some embodiments of the present disclosure, the above
structural unit
##STR00005##
is selected from
##STR00006##
Other variables are as defined by the present disclosure.
[0025] In some embodiments of the present disclosure, the above
structural unit
##STR00007##
is selected from
##STR00008##
Other variables are as defined by the present disclosure.
[0026] In some embodiments of the present disclosure, the above
structural unit
##STR00009##
is selected from
##STR00010##
Other variables are as defined by the present disclosure.
[0027] Some embodiments of the present disclosure are derived from
any combination of the above variables.
[0028] In some embodiments of the present disclosure, the above
compound, the isomer thereof, or the pharmaceutically acceptable
salt thereof is selected from the group consisting of
##STR00011##
[0029] wherein,
[0030] R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are as defined in the
present disclosure.
[0031] The present disclosure provides a compound of the following
formula, an isomer thereof, or a pharmaceutically acceptable salt
thereof, selected from
##STR00012## ##STR00013##
[0032] The present disclosure provides a pharmaceutical
composition, comprising a therapeutically effective amount of the
above compound, the isomer thereof or the pharmaceutically
acceptable salt thereof as an active ingredient, and a
pharmaceutically acceptable carrier.
[0033] The present disclosure provides use of the above compound or
the pharmaceutically acceptable salt thereof or the above
pharmaceutical composition in the manufacture of a medicament for
treating SGLT1/SGLT2 related diseases.
[0034] In some embodiments of the present disclosure, the disease
is diabetes.
Technical Effect
[0035] The compound of the present disclosure exhibits superior
inhibitory activity against Human-SGLT1 and Human-SGLT2 in vitro,
and exhibits good hypoglycemic effect in animals.
Definition and Description
[0036] Unless otherwise stated, the following terms and phrases as
used herein are intended to have the following meanings. A
particular term or phrase should not be considered undefined or
unclear without a particular definition, but should be understood
in the ordinary sense. When a trade name appears herein, it is
intended to refer to its corresponding commodity or its active
ingredient. The term "pharmaceutically acceptable" as used herein
is intended to mean that those compounds, materials, compositions
and/or dosage forms are within the scope of sound medical judgment
and are suitable for use in contact with human and animal tissues
without excessive toxicity, irritation, allergic reactions or other
problems or complications, and commensurate with a reasonable
benefit/risk ratio.
[0037] The term "pharmaceutically acceptable salt" refers to a salt
of the compound of the present disclosure, prepared from a compound
having a particular substituent found in the present disclosure and
a relatively non-toxic acid or base. When a relatively acidic
functional group is contained in the compound of the present
disclosure, a base addition salt can be obtained by contacting a
neutral form of such a compound with a sufficient amount of a base
in a pure solution or a suitable inert solvent. Pharmaceutically
acceptable base addition salts include sodium salts, potassium
salts, calcium salts, ammonium salts, organic amine salts or
magnesium salts or similar salts. When a relatively basic
functional group is contained in the compound of the present
disclosure, an acid addition salt can be obtained by contacting a
neutral form of such a compound with a sufficient amount of an acid
in a pure solution or a suitable inert solvent. Examples of
pharmaceutically acceptable acid addition salts include inorganic
acid salts, wherein the inorganic acid includes, for example,
hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid,
hydrogen carbonate, phosphoric acid, monohydrogen phosphate,
dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic
acid, and phosphorous acid; and organic acid salts, wherein the
organic acid includes, for example, 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, and methanesulfonic acid; and further include
salts of amino acids (such as arginine, etc.), and salts of organic
acids such as glucuronic acid. Certain specific compounds of the
present disclosure contain both basic and acidic functional groups
and thus can be converted to any base or acid addition salts.
[0038] The pharmaceutically acceptable salts of the present
disclosure can be synthesized from the parent compound containing
an acid group or a base by conventional chemical methods. In
general, such salts are prepared by reacting these compounds via a
free acid or base form with a stoichiometric amount of the
appropriate base or acid in water or an organic solvent or a
mixture of the two.
[0039] In addition to forms of salts thereof, the compounds
provided by the present disclosure also exist in forms of prodrug
thereof. The prodrugs of the compounds described herein easily
chemically transformed under physiological conditions to convert
into the compound of the present disclosure. In addition, the
prodrug may be converted to the compounds of the present disclosure
by chemical or biochemical methods in the in vivo environment.
[0040] Certain compounds of the present disclosure may exist in
unsolvated or solvated forms, including hydrated forms. Generally,
the solvated forms and the unsolvated forms are both included in
the scope of the present disclosure.
[0041] The compounds of the present disclosure may exist in
specific geometric or stereoisomeric forms. The present disclosure
contemplates all such compounds, including the cis and trans
isomers, the (-)- and (+)-enantiomers, the (R)- and
(S)-enantiomers, and the diastereoisomer, a (D)-isomer, a
(L)-isomer, and a racemic mixture thereof, and other mixtures such
as the mixtures enriched in enantiomer and diastereoisomer, all of
which are within the scope of the present disclosure. Additional
asymmetric carbon atoms may be present in the substituents such as
alkyl groups. All such isomers as well as mixtures thereof are
included within the scope of the present disclosure.
[0042] Unless otherwise indicated, the term "enantiomer" or
"optical isomer" refers to a stereoisomer that is a mirror image of
each other.
[0043] Unless otherwise indicated, the terms "cis-trans isomer" or
"geometric isomer" are caused by the fact that double bonds or
single bonds of ring-forming carbon atoms cannot rotate freely.
[0044] Unless otherwise indicated, the term "diastereomer" refers
to a stereoisomer in which a molecule has two or more chiral
centers and a nonmirror image relationship exists between the
molecules.
[0045] Unless otherwise indicated, "(D)" or "(+)" means
dextrorotation, "(L)" or "(-)" means levorotation, "(DL)" or
"(.+-.)" means racemization.
[0046] Unless otherwise stated, the solid wedge bond () and the
wedge dashed bond () represent the absolute configuration of a
stereocenter, the straight solid bond () and the straight dashed
bond () represent the relative configuration of a stereocenter, the
wavy line () represents the solid wedge bond () or the wedge dashed
bond (), the wavy line () represents the straight solid bond () and
the straight dashed bond) ().
[0047] The compounds of the present disclosure may be present in
particular. Unless otherwise indicated, the terms "tautomer" or
"tautomeric form" mean that the different functional isomers are in
dynamic equilibrium at room temperature and can be rapidly
converted into each other. If tautomers are possible (such as in a
solution), the chemical equilibrium of the tautomers can be
achieved. For example, proton tautomer (also known as prototropic
tautomer) includes interconversions by proton transfer, such as
keto-enol isomerization and imine-enamine isomerization. The
valence tautomer includes mutual transformation through the
recombination of some of the bonding electrons. A specific example
of keto-enol tautomerization is the interconversion between two
tautomers pentane-2,4-dione and 4-hydroxypent-3-en-2-one.
[0048] Unless otherwise indicated, the terms "Being rich in one
isomer", "isomer enriched", "being rich in one enantiomer" or
"enantiomer enriched" refer to the content of one of the isomers or
enantiomers is smaller than 100%, and the content of the isomer or
enantiomer is greater than or equal to 60%, or greater than or
equal to 70%, or greater than or equal to 80%, or greater than or
equal to 90%, or greater than or equal to 95%, or greater than or
equal to 96%, or greater than or equal to 97%, or greater than or
equal to 98%, or greater than or equal to 99%, or greater than or
equal to 99.5%, or greater than or equal to 99.6%, or greater than
or equal to 99.7%, or greater than or equal to 99.8%, or greater
than or equal to 99.9%.
[0049] Unless otherwise indicated, the term "isomer excess" or
"enantiomeric excess" refers to the relative percentage difference
between two isomers or two enantiomers. For example, if one of the
isomers or enantiomers is present in an amount of 90% and the other
isomer or enantiomer is present in an amount of 10%, the isomer or
enantiomeric excess (ee value) is 80%.
[0050] The optically active (R)- and (S)-isomers as well as the D
and L isomers can be prepared by chiral synthesis or chiral
reagents or other conventional techniques. If an enantiomer of the
compound of the present disclosure is desired, it can be prepared
by asymmetric synthesis or by derivatization with a chiral
auxiliary, wherein the resulting mixture of diastereomers is
separated and the auxiliary group is cleaved to provide the desired
pure enantiomer. Alternatively, when a molecule contains a basic
functional group (e.g., an amino group) or an acidic functional
group (e.g., a carboxyl group), a diastereomeric salt is formed
with a suitable optically active acid or base, and then the
diastereomers are resolved by conventional methods well known in
the art, and the pure enantiomer is recovered. Furthermore, the
separation of enantiomers and diastereomers is generally
accomplished by the use of chromatography using a chiral stationary
phase optionally combined with chemical derivatization (eg,
formation of carbamate from an amine). The compounds of the present
disclosure may contain an unnatural proportion of atomic isotopes
on one or more of the atoms constituting the compound. For example,
a compound can be labeled with a radioisotope such as tritium
(.sup.3H), iodine-125 (.sup.125I) or C-14 (.sup.14C). For another
example, the hydrogen can be replaced by heavy hydrogen to form
deuterated drugs. The bond formed by deuterium and carbon is
stronger than the bond formed by ordinary hydrogen and carbon.
Compared with undeuterated drugs, the deuterated drugs have
advantages such as reduced toxic and side effects, an increased
stability, a strengthen efficacy, and prolonged biological
half-life. Alterations of all isotopes of the compounds of the
present disclosure, whether radioactive or not, are included within
the scope of the present disclosure. The term "pharmaceutically
acceptable carrier" refers to any formulation or carrier medium
that is capable of delivering an effective amount of an active
substance of the present disclosure, does not interfere with the
biological activity of the active substance, and has no toxic and
side effects on the host or patient, and the representative carrier
includes water, oil, vegetables and minerals, cream bases, lotion
bases, ointment bases, etc. These bases include suspending agents,
tackifiers, transdermal enhancers and the like. Their formulations
are well known to those skilled in the cosmetic or topical
pharmaceutical arts.
[0051] The term "excipient" generally refers to a carrier, a
diluent and/or a medium required to prepare an effective
pharmaceutical composition.
[0052] The term "effective amount" or "therapeutically effective
amount", with respect to a pharmaceutical or pharmacologically
active agent, refers to a sufficient amount of a drug or agent that
is non-toxic but that achieves the desired effect. For oral dosage
forms in the present disclosure, an "effective amount" of an active
substance in a composition refers to the amount required to achieve
the desired effect when used in combination with another active
substance in the composition. The determination of the effective
amount will vary from person to person, depending on the age and
general condition of the recipient, and also depending on the
particular active substance, and a suitable effective amount in an
individual case can be determined by one skilled in the art based
on routine experimentation.
[0053] The term "active ingredient", "therapeutic agent", "active
substance" or "active agent" refers to a chemical entity that is
effective in treating a target disorder, disease or condition.
[0054] "Optional" or "optionally" means that the subsequently
described event or condition may, but is not necessarily, occur,
and the description includes instances in which the event or
condition occurs and instances in which the event or condition does
not occur.
[0055] The term "substituted" means that any one or more hydrogen
atoms on a particular atom are replaced by a substituent, and may
include variants of heavy hydrogen and hydrogen, as long as the
valence of the particular atom is normal and the substituted
compound is stable. When the substituent is a keto group (ie,
.dbd.O), it means that two hydrogen atoms are substituted. Keto
substitution does not occur on the aryl group. The term "optionally
substituted" means that it may or may not be substituted, and
unless otherwise specified, the type and number of substituents may
be arbitrary as long as it is chemically achievable.
[0056] When any variable (eg, R) occurs one or more times in the
composition or structure of a compound, its definition in each case
is independent. Thus, for example, if a group is substituted by 0-2
R groups, the group may optionally be substituted at most by two R
groups, and R has an independent option in each case. Furthermore,
combinations of substituents and/or variants thereof are
permissible only if such combinations result in stable
compounds.
[0057] When the number of one linking group is 0, such as
--(CRR).sub.0--, it indicates that the linking group is a single
bond.
[0058] When one of the variables is selected from a single bond, it
means that the two groups to which it is attached are directly
linked. For example, when L represents a single bond in A-L-Z, the
structure is actually A-Z.
[0059] When a substituent is vacant, it means that the substituent
is absent. For example, when X is vacant in A-X, the structure is
actually A. When the substituents listed do not indicate which atom
is attached to the substituted group, such a substituent may be
bonded through any atom thereof. For example, as a substituent,
pyridyl can be attached to the substituted group through any carbon
atom in the pyridine ring.
[0060] When the listed linking group does not indicate its
attachment direction, its attachment direction is arbitrary. For
example, the linking group L in
##STR00014##
is -M-W-, and at this time -M-W- may connect the ring A and ring B
to form
##STR00015##
according to the direction the same as the reading direction of
from left to right, or -M-W- may connect the ring A and ring B to
form
##STR00016##
according to the direction opposite to the reading direction of
from left to right. The combination of the linking groups,
substituents and/or variants thereof is permitted only if such
combination produces a stable compound.
[0061] Unless otherwise specified, the term "hetero" denotes a
heteroatom or a heteroatom group (ie, a radical containing a
heteroatom), including atoms other than carbon (C) and hydrogen
(H), and radicals containing such heteroatoms, including, for
example, oxygen (O), nitrogen (N), sulfur (S), silicon (Si),
germanium (Ge), aluminum (Al), boron (B), --O--, --S--, .dbd.O,
.dbd.S, --C(.dbd.O)O--, --C(.dbd.O)--, --C(.dbd.S)--, --S(.dbd.O),
--S(.dbd.O).sub.2--, and optionally substituted --C(.dbd.O)N(H)--,
--N(H)--, --C(.dbd.NH)--, --S(.dbd.O).sub.2N(H)-- or
--S(.dbd.O)N(H)--.
[0062] Unless otherwise specified, the term "ring" means a
substituted or unsubstituted cycloalkyl, heterocycloalkyl,
cycloalkenyl, heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl,
aryl, or heteroaryl. The ring includes monocyclic ring, and also
includes bicyclic or polycyclic systems, wherein the bicyclic
system includes spiro ring, fused ring, and bridge ring. The number
of atoms on a ring is usually defined as the number of members of
the ring. For example, a "5-7 membered ring" means 5 to 7 atoms are
arranged in a circle. Unless otherwise specified, the ring
optionally contains 1 to 3 heteroatoms. Thus, the "5-7 membered
ring" includes, for example, phenyl, pyridyl, and piperidyl; on the
other hand, the term "5-7 membered heterocycloalkyl" includes
pyridyl and piperidyl, but does not include phenyl. The term "ring"
also includes a ring system containing at least one ring, wherein
each "ring" independently conforms to the above definition.
[0063] Unless otherwise specified, the term "alkyl" represents a
linear or branched saturated hydrocarbon group. In some
embodiments, the alkyl is C.sub.1-12 alkyl; in other embodiments,
the alkyl is C.sub.1-6 alkyl; in other embodiments, the alkyl is
C.sub.1-3 alkyl. The alkyl can be monosubstituted (eg, --CH.sub.2F)
or polysubstituted (eg, --CF.sub.3), and may be monovalent (eg,
methyl), divalent (such as methylene) or polyvalent (such as
methine). Examples of the alkyl include, but are not limited to,
methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl),
butyl (including n-butyl, isobutyl, s-butyl, t-butyl), pentyl
(including n-pentyl, isopentyl, neopentyl) and the like.
[0064] Unless otherwise specified, the term "alkenyl" represents a
linear or branched hydrocarbon group containing one or more
carbon-carbon double bonds located at any position of the group. In
some embodiments, the alkenyl is C.sub.2-8 alkenyl; in other
embodiments, the alkenyl is C.sub.2-6 alkenyl; in other
embodiments, the alkenyl is C.sub.2-4 alkenyl. The alkenyl may be
monosubstituted or polysubstituted, and may be monovalent, divalent
or polyvalent. Examples of the alkenyl include, but are not limited
to, vinyl, propenyl, butenyl, pentenyl, hexenyl, butadienyl,
piperylene, hexadienyl and the like.
[0065] Unless otherwise specified, the term "alkynyl" represents a
linear or branched hydrocarbon group containing one or more
carbon-carbon triple bonds located at any position of the group. In
some embodiments, the alkynyl is C.sub.2-8 alkynyl; in other
embodiments, the alkynyl is C.sub.2-6 alkynyl; in other
embodiments, the alkynyl is C.sub.2-4 alkynyl. The alkynyl may be
monosubstituted or polysubstituted, and may be monovalent, divalent
or polyvalent. Examples of the alkynyl include, but are not limited
to, ethynyl, propynyl, butynyl, pentynyl and the like.
[0066] Unless otherwise specified, the term "heteroalkyl" by itself
or in combination with another term refers to a stable linear or
branched alkyl radical or a combination thereof having a certain
number of carbon atoms and at least one heteroatom or heteroatom
group. In some embodiments, the heteroatom is selected from B, O,
N, and S, wherein the nitrogen and sulfur atoms are optionally
oxidized, and the nitrogen heteroatoms are optionally quaternized.
In other embodiments, the heteroatom group is selected from
--C(.dbd.O)O--, --C(.dbd.O)--, --C(.dbd.S)--, --S(.dbd.O),
--S(.dbd.O).sub.2--, --C(.dbd.O)N(H)--, --N(H)--, --C(.dbd.NH)--,
--S(.dbd.O).sub.2N(H)-- and --S(.dbd.O)N(H)--. In some embodiments,
the heteroalkyl is C.sub.1-6 heteroalkyl; in other embodiments, the
heteroalkyl is C.sub.1-3 heteroalkyl. The heteroatom or heteroatom
group may be located at any internal position of the heteroalkyl,
including a position where the alkyl is attached to the rest of the
molecule. The terms "alkoxyl", "alkylamino" and "alkylthio" (or
thioalkoxy) belong to a customary expression, and refer to those
alkyl groups which are attached to the remainder of the molecule
through an oxygen atom, an amino group or a sulfur atom,
respectively. Examples of the heteroalkyl include, but are not
limited to: --OCH.sub.3, --OCH.sub.2CH.sub.3,
--OCH.sub.2CH.sub.2CH.sub.3, --OCH.sub.2(CH.sub.3).sub.2,
--CH.sub.2--CH.sub.2--O--CH.sub.3, --NHCH.sub.3,
--N(CH.sub.3).sub.2, --NHCH.sub.2CH.sub.3,
--N(CH.sub.3)(CH.sub.2CH.sub.3),
--CH.sub.2--CH.sub.2--NH--CH.sub.3,
--CH.sub.2--CH.sub.2--N(CH.sub.3)--CH.sub.3, --SCH.sub.3,
--SCH.sub.2CH.sub.3, --SCH.sub.2CH.sub.2CH.sub.3,
--SCH.sub.2(CH.sub.3).sub.2, --CH.sub.2--S--CH.sub.2--CH.sub.3,
--CH.sub.2--CH.sub.2, --S(.dbd.O)--CH.sub.3,
--CH.sub.2--CH.sub.2--S(.dbd.O).sub.2--CH.sub.3,
--CH.dbd.CH--O--CH.sub.3, --CH.sub.2--CH.dbd.N--OCH.sub.3 and
--CH.dbd.CH--N(CH.sub.3)--CH.sub.3. At most two heteroatoms can be
continuous, such as --CH.sub.2--NH--OCH.sub.3.
[0067] Unless otherwise specified, the term "heteroalkenyl" by
itself or in combination with another term refers to a stable
linear or branched alkenyl radical or a combination thereof having
a certain number of carbon atoms and at least one heteroatom or
heteroatom group. In some embodiments, the heteroatom is selected
from B, O, N, and S, wherein the nitrogen and sulfur atoms are
optionally oxidized, and the nitrogen heteroatoms are optionally
quaternized. In other embodiments, the heteroatom group is selected
from --C(.dbd.O)O--, --C(.dbd.O)--, --C(.dbd.S)--, --S(.dbd.O),
--S(.dbd.O).sub.2--, --C(.dbd.O)N(H)--, --N(H)--, --C(.dbd.NH)--,
--S(.dbd.O).sub.2N(H)-- and --S(.dbd.O)N(H)--. In some embodiments,
the heteroalkenyl is C.sub.2-6 heteroalkenyl; in other embodiments,
the heteroalkyl is C.sub.2-4 heteroalkenyl. The heteroatom or
heteroatom group may be located at any internal position of the
heteroalkenyl, including a position where the alkenyl is attached
to the rest of the molecule. The terms "alkenyloxy", "alkenylamino"
and "alkenylthio" belong to a customary expression, and refer to
those alkenyl groups which are attached to the remainder of the
molecule through an oxygen atom, an amino group or a sulfur atom,
respectively. Examples of the heteroalkenyl include, but are not
limited to, --O--CH.dbd.CH.sub.2, --O--CH.dbd.CHCH.sub.3,
--O--CH.dbd.C(CH.sub.3).sub.2, --CH.dbd.CH--O--CH.sub.3,
--O--CH.dbd.CHCH.sub.2CH.sub.3, --CH.sub.2--CH.dbd.CH--OCH.sub.3,
--NH--CH.dbd.CH.sub.2, --N(CH.dbd.CH.sub.2)--CH.sub.3,
--CH.dbd.CH--NH--CH.sub.3, --CH.dbd.CH--N(CH.sub.3).sub.2,
--S--CH.dbd.CH.sub.2, --S--CH.dbd.CHCH.sub.3,
--S--CH.dbd.C(CH.sub.3).sub.2, --CH.sub.2--S--CH.dbd.CH.sub.2,
--S(.dbd.O)--CH.dbd.CH.sub.2 and
--CH.dbd.CH--S(.dbd.O).sub.2--CH.sub.3. At most two heteroatoms can
be continuous, such as --CH.dbd.CH--NH--OCH.sub.3.
[0068] Unless otherwise specified, the term "heteroalkynyl" by
itself or in combination with another term refers to a stable
linear or branched alkynyl radical or a combination thereof having
a certain number of carbon atoms and at least one heteroatom or
heteroatom group. In some embodiments, the heteroatom is selected
from B, O, N, and S, wherein the nitrogen and sulfur atoms are
optionally oxidized, and the nitrogen heteroatom is optionally
quaternized. In other embodiments, the heteroatom group is selected
from --C(.dbd.O)O--, --C(.dbd.O)--, --C(.dbd.S)--, --S(.dbd.O),
--S(.dbd.O).sub.2--, --C(.dbd.O)N(H)--, --N(H)--, --C(.dbd.NH)--,
--S(.dbd.O).sub.2N(H)-- and --S(.dbd.O)N(H)--. In some embodiments,
the heteroalkynyl is C.sub.2-6 heteroalkynyl; in other embodiments,
the heteroalkyl is C.sub.2-4 heteroalkynyl. The heteroatom or
heteroatom group may be located at any internal position of the
heteroalkynyl, including a position where the alkynyl is attached
to the rest of the molecule. The terms "alkynyloxy", "alkynylamino"
and "alkynylthio" belong to a customary expression, and refers to
those alkynyl groups which are attached to the remainder of the
molecule through an oxygen atom, an amino group, or a sulfur atom,
respectively. Examples of the heteroalkynyl include, but are not
limited to
##STR00017##
At most two heteroatoms can be continuous, such as
##STR00018##
[0069] Unless otherwise specified, the term "cycloalkyl" includes
any stable cyclic alkyl group, including monocyclic, bicyclic, or
tricyclic systems, wherein the bicyclic and tricyclic systems
include a spiro ring, a fused ring, and a bridge ring. In some
embodiments, the cycloalkyl is C.sub.3-8 cycloalkyl; in other
embodiments, the cycloalkyl is C.sub.3-6 cycloalkyl; in other
embodiments, the cycloalkyl is C.sub.5-6 cycloalkyl. The cycloalkyl
may be monosubstituted or polysubstituted, and may be monovalent,
divalent or polyvalent. Examples of these cycloalkyl groups
include, but are not limited to, cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, cycloheptyl, norbornyl,
[2.2.2]bicyclooctane, [4.4.0]bicyclodecane and the like.
[0070] Unless otherwise specified, the term "cycloalkenyl" includes
any stable cyclic alkenyl group containing one or more unsaturated
carbon-carbon double bonds at any position of the group, which
includes monocyclic, bicyclic or tricyclic systems, wherein the
bicyclic and tricyclic systems include a spiro ring, a fused ring,
and a bridge ring, but any ring in this system is non-aromatic. In
some embodiments, the cycloalkenyl is C.sub.3-8 cycloalkenyl; in
other embodiments, the cycloalkenyl is C.sub.3-6 cycloalkenyl; in
other embodiments, the cycloalkenyl is C.sub.5-6 cycloalkenyl. The
cycloalkenyl may be monosubstituted or polysubstituted, and may be
monovalent, divalent or polyvalent. Examples of these cycloalkenyl
groups include, but are not limited to, cyclopentenyl, cyclohexenyl
and the like.
[0071] Unless otherwise specified, the term "cycloalkynyl" includes
any stable cyclic alkynyl group containing one or more
carbon-carbon triple bonds at any position of the group, which
includes monocyclic, bicyclic or tricyclic systems, wherein the
bicyclic and tricyclic systems include a spiro ring, a fused ring,
and a bridge ring. It may be monosubstituted or polysubstituted,
and may be monovalent, divalent or polyvalent.
[0072] Unless otherwise specified, the term "heterocycloalkyl" by
itself or in combination with other terms refers to cyclized
"heteroalkyl", which includes monocyclic, bicyclic and tricyclic
systems, wherein the bicyclic and tricyclic systems include a spiro
ring, a fused ring, and a bridge ring. In addition, in the case of
the "heterocycloalkyl", a heteroatom may occupy a position where
the heterocycloalkyl is bonded to the rest of the molecule. In some
embodiments, the heterocycloalkyl is 4-6 membered heterocycloalkyl;
in other embodiments, the heterocycloalkyl is 5-6 membered
heterocycloalkyl. Examples of the heterocycloalkyl include, but are
not limited to, azetidinyl, oxetanyl, thiatanyl, pyrrolidinyl,
pyrazolidinyl, imidazolidinyl, tetrahydrothienyl (including
tetrahydrothiophene-2-yl and tetrahydrothiophen-3-yl, etc.),
tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.),
tetrahydropyranyl, piperidinyl (including 1-piperidinyl,
2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including
1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including
3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl,
isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl,
hexahydropyridazinyl, homopiperazinyl, homopiperidinyl or
oxepanyl.
[0073] Unless otherwise specified, the term "heterocycloalkenyl" by
itself or in combination with other terms refers to cyclized
"heteroalkenyl", which includes monocyclic, bicyclic and tricyclic
systems, wherein the bicyclic and tricyclic systems include a spiro
ring, a fused ring, and a bridge ring, but any ring in this system
is non-aromatic. In addition, in the case of the
"heterocycloalkenyl", a heteroatom may occupy a position where the
heterocycloalkenyl is bonded to the rest of the molecule. In some
embodiments, the heterocycloalkenyl is 4-6 membered
heterocycloalkenyl; in other embodiments, the heterocycloalkenyl is
5-6 membered heterocycloalkenyl. Examples of heterocycloalkenyl
include, but are not limited to,
##STR00019##
[0074] Unless otherwise specified, the term "heterocycloalkynyl" by
itself or in combination with other terms refers to a cyclized
"heteroalkynyl", which includes monocyclic, bicyclic and tricyclic
systems, wherein the bicyclic and tricyclic systems include a spiro
ring, a fused ring, and a bridge ring. In addition, in the case of
the "heterocycloalkynyl", a heteroatom may occupy a position where
the heterocycloalkynyl is bonded to the rest of the molecule. In
some embodiments, the heterocycloalkynyl is 4-6 membered
heterocycloalkynyl; in other embodiments, the heterocycloalkynyl is
5-6 membered heterocycloalkynyl. Unless otherwise specified, the
term "halo" or "halogen", by itself or as a part of another
substituent, refers to a fluorine, chlorine, bromine or iodine
atom. Further, the term "haloalkyl" is intended to include both
monohaloalkyl and polyhaloalkyl. For example, the term
"halo(C.sub.1-C.sub.4)alkyl" is intended to include, but is not
limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl,
3-bromopropyl, etc. Unless otherwise specified, examples of
haloalkyl include, but are not limited to, trifluoromethyl,
trichloromethyl, pentafluoroethyl and pentachloroethyl.
[0075] The term "alkoxyl" represents the above alkyl group having a
specified number of carbon atoms attached through an oxygen bridge,
and unless otherwise specified, C.sub.1-6 alkoxyl includes C.sub.1,
C.sub.2, C.sub.3, C.sub.4, C.sub.5 and C.sub.6 alkoxyl. In some
embodiments, the alkoxy is C.sub.1-3 alkoxy. Examples of alkoxyl
include, but are not limited to, methoxy, ethoxy, n-propoxy,
isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentyloxy and
S-pentyloxy.
[0076] Unless otherwise specified, the terms "aromatic ring" and
"aryl" in the present disclosure may be used interchangeably. The
term "aromatic ring" or "aryl" refers to a polyunsaturated
carbocyclic system, which can be monocyclic, bicyclic or polycyclic
system, wherein at least one ring is aromatic, and the rings are
fused together in the bicyclic and polycyclic systems. It may be
monosubstituted or polysubstituted, and may be monovalent, divalent
or polyvalent. In some embodiments, the aryl is C.sub.6-12 aryl; in
other embodiments, the aryl is C.sub.6-10 aryl. Examples of aryl
include, but are not limited to, phenyl, naphthyl (including
1-naphthyl, 2-naphthyl, etc.). The substituent of any one of the
above aryl ring systems is selected from the acceptable
substituents described in the present disclosure.
[0077] Unless otherwise specified, the terms "heteroaryl ring" and
"heteroaryl" of the present disclosure may be used interchangeably.
The term "heteroaryl" refers to an aryl (or aromatic ring)
containing 1, 2, 3 or 4 heteroatom(s) independently selected from
B, N, O and S. It may be monocyclic, bicyclic or tricyclic systems,
wherein the nitrogen atom may be substituted or unsubstituted (i.e.
N or NR, wherein R is H or other substituents as defined herein)
and optionally quaternized. The nitrogen and sulfur heteroatoms may
be optionally oxidized (i.e. NO and S(O).sub.p, p is 1 or 2).
Heteroaryl may be attached to the remainder of the molecule through
heteroatoms. In some embodiments, the heteroaryl is 5-10 membered
heteroaryl; in other embodiments, the heteroaryl is 5-6 membered
heteroaryl. Examples of the heteroaryl include, but are not limited
to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl,
etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc),
imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and
5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and
5-oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl,
2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl and 4H-1,2,4-triazolyl,
etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl and
5-isoxazolyl, etc), thiazolyl (including 2-thiazolyl, 4-thiazole
and 5-thiazolyl, etc.), furyl (including 2-furyl and 3-furyl,
etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl
(including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl,
pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.),
benzothiazolyl (including 5-benzothiazolyl, etc.), purinyl,
benzimidazolyl (including 2-benzimidazolyl, etc.), indolyl
(including 5-indolyl, etc.), isoquinolinyl (including
1-isoquinolinyl and 5-isoquinolinyl, etc.), quinoxalinyl (including
2-quinoxalinyl and 5-quinoxalinyl, etc.), quinolinyl (including
3-quinolinyl and 6-quinolinyl, etc.), pyrazinyl, purinyl, phenyl
and oxazolyl. The substituent of any one of the above heteroaryl
ring systems is selected from the acceptable substituents described
in the present disclosure.
[0078] Unless otherwise specified, the term "aralkyl" is intended
to include those groups in which an aryl group is attached to an
alkyl group. In some embodiments, the aralkyl is a C.sub.6-10
aryl-C.sub.14 alkyl; in other embodiments, the aralkyl is
C.sub.6-10 aryl-C.sub.1-2 alkyl. Examples of the aralkyl include,
but are not limited to, benzyl, phenethyl, menaphthyl and the like.
The terms "aryloxy" and "arylthio" represent those groups in which
the carbon atom (such as methyl) in the aralkyl has been replaced
by an oxygen or sulfur atom. In some embodiments, the aryloxy is
C.sub.6-10 aryl-O--C.sub.1-2 alkyl; in other embodiments, the
aryloxy is C.sub.6-10 aryl-C.sub.1-2 alkyl-O--. In some
embodiments, the arylthio is C.sub.6-10 aryl-S--C.sub.1-2 alkyl; in
other embodiments, the arylthio is C.sub.6-10 aryl-C.sub.1-2
alkyl-S--. Examples of aryloxy and arylthio include, but are not
limited to, phenoxymethyl, 3-(1-naphthyloxy) propyl,
phenylthiomethyl and the like.
[0079] Unless otherwise specified, the term "heteroaralkyl" is
intended to include those groups in which a heteroaryl group is
attached to an alkyl group. In some embodiments, the heteroaralkyl
is 5-8 membered heteroaryl-C.sub.1-4 alkyl; in other embodiments,
the heteroaralkyl is 5-6 membered heteroalkyl-C.sub.1-2 alkyl.
Examples of heteroaralkyl include, but are not limited to,
pyrrolylmethyl, pyrazolylmethyl, pyridylmethyl, pyrimidinylmethyl
and the like. The terms "heteroaryloxy" and "heteroarylthio"
respectively refer to those groups in which a carbon atom (such as
methyl) in the heteroaralkyl group have been replaced by an oxygen
or sulfur atom. In some embodiments, the heteroaryloxy is 5-8
membered heteroaryl-O--C.sub.1-2 alkyl; in other embodiments, the
heteroaryloxy is 5-6 membered heteroaryl-C.sub.1-2 alkyl-O--. In
some embodiments, the heteroarylthio is 5-8 membered
heteroaryl-S--C.sub.1-2 alkyl; in other embodiments, the
heteroarylthio is 5-6 membered heteroaryl-C.sub.1-2 alkyl-S--.
Examples of heteroaryloxy and heteroarylthio include, but are not
limited to, pyrroleoxymethyl, pyrazolyloxymethyl,
2-pyridyloxymethyl, pyrrolylthiomethyl, pyrazolylthiomethyl,
2-pyridylthiomethyl and the like.
[0080] Unless otherwise specified, C.sub.n-n+m or
C.sub.n--C.sub.n+m includes cases where the carbon number is a
specific number from n to n+m, and also includes cases wherein the
carbon number is in a range within n to n+m. For example,
C.sub.1-12 includes C.sub.1, C.sub.2, C.sub.3, C.sub.4, C.sub.5,
C.sub.6, C.sub.7, C.sub.8, C.sub.9, C.sub.10, C.sub.11 and
C.sub.12, and also includes C.sub.1-3, C.sub.1-6, C.sub.1-9,
C.sub.3-6, C.sub.3-9, C.sub.3-12, C.sub.6-9, C.sub.6-12 and
C.sub.9-12, etc. Similarly, n to n+m membered ring means that the
number of atoms in the ring is from n to n+m, or the number of
atoms is in any range from n to n+m. For example, 3-12 membered
ring includes 3 membered ring, 4 membered ring, 5 membered ring, 6
membered ring, 7 membered ring, 8 membered ring, 9 membered ring,
10 membered ring, 11 membered ring and 12 membered ring, and also
includes 3-6 membered ring, 3-9 membered ring, 5-6 membered ring,
5-7 membered ring, 6-7 membered ring, 6-8 membered ring, and 6-10
membered ring.
[0081] The term "leaving group" refers to a functional group or
atom which may be substituted by another functional group or atom
by a substitution reaction (for example, a nucleophilic
substitution reaction). For example, representative leaving groups
include triflate; chlorine, bromine, and iodine; sulfonate groups
such as methanesulfonate, tosylate, p-bromobenzene sulfonate,
p-tosylate and the like; acyloxyl such as acetoxyl,
trifluoroacetoxyl and the like.
[0082] The term "protecting group" includes but is not limited to
"amino protecting group", "hydroxy protecting group" or "mercapto
protecting group". The term "amino protecting group" refers to a
protecting group suitable for preventing side reactions at the
nitrogen position of the amino group. Representative amino
protecting groups include, but are not limited to, formyl, acyl
such as alkanoyl (such as acetyl, trichloroacetyl, or
trifluoroacetyl), alkoxycarbonyl such as t-butoxycarbonyl (Boc),
arylmethoxycarbonyl such as carbobenzoxy (Cbz) and
9-fluorenylmethoxycarbonyl (Fmoc), arylmethyl such as benzyl (Bn),
trityl (Tr), 1,1-di-(4'-methoxyphenyl) methyl, and silyl such as
trimethylsilyl (TMS) and t-butyldimethylsilyl (TBS), etc. The term
"hydroxyl protecting group" refers to a protecting group suitable
for preventing side reactions of hydroxyl groups. Representative
hydroxy protecting groups include, but are not limited to, alkyl
such as methyl, ethyl, and t-butyl, acyl such as alkanoyl (such as
acetyl), arylmethyl such as benzyl (Bn), p-methyl Oxybenzyl (PMB),
9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM),
and silyl such as trimethylsilyl (TMS) and t-butyldimethylsilyl
(TBS), etc.
[0083] The compounds of the present disclosure may be prepared by a
variety of synthetic methods well known to those skilled in the
art, including the specific embodiments set forth below,
embodiments formed through combinations thereof with other chemical
synthetic methods, and those equivalent alternatives well known to
those skilled in the art, and preferred embodiments include, but
are not limited to, embodiments of the present disclosure.
[0084] The compounds of the present disclosure can have various
applications or can be used to treat various diseases, including
but not limited to the specific applications or diseases listed
herein.
[0085] The solvent used in the present disclosure is commercially
available. The present disclosure employs the following
abbreviations: aq stands for water; HATU stands for
O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyl urea
hexafluorophosphate; EDC stands for
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride;
m-CPBA stands for 3-chloroperoxybenzoic acid; eq stands for
equivalent weight, and equal weight; CDI stands for carbonyl
diimidazole; DCM stands for dichloromethane; PE stands for
petroleum ether; DIAD stands for diisopropyl azodicarboxylate; DMF
stands for N,N-dimethylformamide; DMSO stands for dimethyl
sulfoxide; EtOAc represents ethyl acetate; EtOH represents ethanol;
MeOH represents methanol; CBz represents benzyloxycarbonyl, an
amine protecting group; BOC represents t-butoxycarbonyl which is an
amine protecting group; HOAc represents acetic acid; NaCNBH.sub.3
represents sodium cyanoborohydride; r.t. represents room
temperature; O/N stands for overnight; THF stands for
tetrahydrofuran; Boc.sub.2O stands for di-tert-butyldicarbonate;
TFA stands for trifluoroacetic acid; DIPEA stands for
diisopropylethylamine; SOCl.sub.2 stands for thionyl chloride;
CS.sub.2 stands for carbon disulfide; TsOH stands for
p-toluenesulfonic acid; NFSI stands for
N-fluoro-N-(phenylsulfonyl)benzenesulfonamide; NCS stands for
N-chlorosuccinimide; n-Bu.sub.4NF stands for tetrabutylammonium
fluoride; iPrOH stands for 2-propanol; mp stands for melting point;
LDA stands for lithium diisopropylamide; and NMP stands for
N-methyl pyrrolidone.
[0086] Compounds are named by hand or by ChemDraw.RTM. software,
and commercial compounds are based on supplier catalog names.
BRIEF DESCRIPTION OF THE DRAWINGS
[0087] FIG. 1: Results of changes in the body weight of animal from
week 1 to week 8;
[0088] FIG. 2: Results of changes in the food consumption of animal
from week 1 to week 8.
DETAILED DESCRIPTION
[0089] The present disclosure is described in detail below by
referring to the examples, which are not intended to adversely
limit the present disclosure. The present disclosure has been
described in detail herein, the embodiments of the present
disclosure are disclosed herein, and various modifications and
changes may be made to the embodiments of the present disclosure
without departing from the spirit and scope of the present
disclosure, which is obvious for those skilled in the art.
Reference Example 1: Fragment A-1
##STR00020##
[0090] Synthesis Route:
##STR00021##
[0092] Step 1: Synthesis of Compound A-1-3
[0093] Compound A-1-1 (20 g, 84.78 mmol, 10.87 mL, 1 eq) and
tetrahydrofuran (125 mL) were successively added to a pre-dried
three-necked flask (500 mL). After replaced with nitrogen, the
flask was cooled to -78.degree. C., and N-butyllithium (2.5 M,
37.64 mL, 1.11 eq) was slowly added dropwise thereto, and then
stirring was performed for 0.5 hours. Finally, compound A-1-2 (12.5
g, 93.26 mmol, 1.1 eq) was added to the flask, and then the
temperature was slowly raised to 0.degree. C., and stirring was
performed for 0.5 hours. After the reaction was completed, the
resulted reaction solution was slowly quenched with a saturated
aqueous solution of ammonium chloride (200 mL) at 0-10.degree. C.
Then the reaction solution was extracted with ethyl acetate (200
mL.times.2) to obtain organic phases. The combined organic phases
were washed with saturated sodium chloride (100 mL), dried with
anhydrous sodium sulfate desiccant, filtered to remove the
desiccant, and concentrated in vacuo to remove the solvent,
obtaining a crude compound A-1-3, which was directly used in the
next reaction without purification.
[0094] Step 2: Synthesis of Compound A-1-4.
[0095] Compound A-1-3 (23.2 g, 79.82 mmol, 1 eq) and toluene (600
mL) were successively added to a pre-dried three-necked flask (1000
mL), and then p-toluenesulfonic acid monohydrate (1.82 g, 9.58
mmol, 0.12 eq) was added thereto. After replaced with nitrogen, the
flask was heated to 130.degree. C., and stirring was performed for
10 hours (equipped with Dean-Stark). After the reaction was
completed, the reaction solution was cooled down, and concentrated
in vacuo to remove the solvent to obtain a residue. The residue was
subjected to column chromatography (petroleum ether/ethyl acetate
system) to separate compound A-1-4. .sup.1H NMR (400 MHz,
CHLOROFORM-d) .delta.: 7.49-7.43 (m, 2H), 7.27-7.22 (m, 2H), 5.91
(dt, J=1.3, 2.6 Hz, 1H), 2.80-2.63 (m, 4H), 2.19 (tt, J=6.7, 13.7
Hz, 2H).
[0096] Step 3: Synthesis of Compound A-1.
[0097] Compound A-1-4 (2.9 g, 10.62 mmol, 1 eq), pinacol borate
(5.39 g, 21.24 mmol, 2 eq), potassium acetate (3.13 g, 31.85 mmol,
3 eq) and 1,4-dioxane (30 mL) were successively added to a
pre-dried one-necked 100 mL flask. After replaced with nitrogen,
1,1'-bis(diphenylphosphino)ferrocene palladium chloride (776.94 mg,
1.06 mmol, 0.1 eq) was added into the reaction. After replaced with
nitrogen again, the flask was heated to 70.degree. C. and stirring
was performed for 10 hours. After the reaction was completed, the
reaction was cooled down and concentrated in vacuo to remove the
solvent to obtain a residue. The residue was purified by column
chromatography (petroleum ether/ethyl acetate system) to obtain
compound A-1. .sup.1H NMR (400 MHz, CHLOROFORM-d) .delta.: 7.78 (d,
J=8.4 Hz, 2H), 7.38 (d, J=8.2 Hz, 2H), 5.97 (br s, 1H), 2.79-2.66
(m, 4H), 2.19 (tt, J=6.6, 13.7 Hz, 2H), 1.36 (s, 12H).
[0098] The fragments A2-8 in the following table are synthesized
with reference to the steps 1-3 as described in the Reference
Example 1. The structures in the table also include their possible
isomers.
TABLE-US-00001 Reference Fragments Example A Structure .sup.1H NMR
2 A-2 ##STR00022## .sup.1H NMR (400 M Hz, CHLOROFORM-d) .delta. ppm
1.35 (s, 12H), 2.49-2.60 (m, 2H), 3.94 (t, J = 5.40 Hz, 2H), 4.33
(q, J = 2.76 Hz, 2H), 6.19 (dt, J = 2.82, 1.47 Hz, 1H), 7.40 (d, J
= 8.03 Hz, 2H), 7.76-7.81 (m, 2H) 3 A-3 ##STR00023## .sup.1H NMR
(400 M Hz, CHLOROFORM-d) .delta. ppm 7.75 (d, J = 8.3 Hz, 2H), 7.40
(d, J = 8.0 Hz, 2H), 6.03-6.07 (m, 1H), 4.03 (s, 4H), 2.64-2.72 (m,
2H), 2.48 (br d, J = 1.3 Hz, 2H), 1.93 (t, J = 6.4 Hz, 2H), 1.35
(s, 12H) 4 A-4 ##STR00024## .sup.1H NMR (400 M Hz, CHLOROFORM-d)
.delta. ppm 1.35 (s, 12H), 1.62-1.72 (m, 2H), 1.74-1.85 (m, 2H),
2.18-2.26 (m, 2H), 2.37-2.46 (m, 2H), 6.19 (dt, J = 3.76, 2.13 Hz,
1H), 7.39 (d, J = 8.28 Hz, 2H), 7.76 (d, J = 8.03 Hz, 2H) 5 A-5
##STR00025## .sup.1H NMR (400 M Hz, CHLOROFORM-d) .delta. ppm 7.75
(d, J = 8.1 Hz, 2H), 7.43 (d, J = 8.1 Hz, 2H), 6.26 (t, J = 1.9 Hz,
1H), 2.68-2.76 (m, 2H), 2.54 (td, J = 7.4, 2.4 Hz, 2H), 1.98-2.05
(m, 2H), 1.35 (s, 12H) 6 A-6 ##STR00026## .sup.1H NMR (400 M Hz,
CHLOROFORM-d) .delta. ppm 7.74 (d, J = 8.0 Hz, 2H), 7.32 (d, J =
8.0 Hz, 2H), 6.14 (t, J = 6.8 Hz, 1H), 2.57-2.66 (m, 2H), 2.27-
2.33 (m, 2H), 1.81-1.87 (m, 2H), 1.65 (dt, J = 11.0, 5.6 Hz, 2H),
1.57 (s, 2H), 1.34-1.36 (m, 12H) 7 A-7 ##STR00027## .sup.1H NMR
(400 M Hz, CHLOROFORM-d) .delta. ppm 7.43 (d, J = 3.5 Hz, 1H),
6.98-7.00 (m, 1H), 6.00 (br s, 1H), 2.79-2.69 (m, 4H), 2.03-2.18
(m, 2H), 1.27 (s, 12H) 8 A-8 ##STR00028## .sup.1H NMR (400 M Hz,
CHLOROFORM-d) .delta. ppm 1.35 (s, 12H), 2.46-2.64 (m, 2H), 3.92
(t, J = 5.52 Hz, 2H), 4.30 (q, J = 2.76 Hz, 2H), 6.21 (dt, J =
3.01, 1.51 Hz, 1H), 7.07 (d, J = 3.51 Hz, 1H), 7.52 (d, J = 3.51
Hz, 1H)
Reference Example 9: Fragment B-1
##STR00029##
[0100] Synthesis Route:
##STR00030## ##STR00031##
[0101] Step 1: Synthesis of Compound B-1-2
[0102] Compound B-1-1 (30 g, 127.41 mmol, 1 eq) and tetrahydrofuran
(6 mL) were added to a 3 L three-necked flask, and the borane
tetrahydrofuran complex (1 M, 382.23 mL, 3 eq) was added thereto
under blowing nitrogen protection. The resulted mixture was reacted
at 25.degree. C. for 16 hours. After the reaction was completed,
the resulted reaction solution was quenched by dropwise addition
with methanol (150 mL) at 25.degree. C. under blowing nitrogen
protection. Then the reaction solution was concentrated in vacuo at
45.degree. C. with a water pump to obtain compound B-1-2. .sup.1H
NMR (400 MHz, CHLOROFORM-d) .delta.=7.68 (d, J=2.4 Hz, 1H), 7.37
(dd, J=2.2, 8.6 Hz, 1H), 7.22 (d, J=8.4 Hz, 1H), 4.77 (d, J=5.3 Hz,
2H).
[0103] Step 2: Synthesis of Compound B-1-3.
[0104] Compound B-1-2 (27 g, 121.91 mmol, 1 eq) and
dimethylformamide (150 mL) were added to a three-necked flask.
After protected with nitrogen, sodium hydride (9.75 g, 243.82 mmol,
60% purity, 2 eq) was added thereto at 0.degree. C. Half an hour
later, allyl bromide (44.24 g, 365.73 mmol, 32.06 mL, 3 eq) was
added thereto, and the resulted mixture was reacted at 25.degree.
C. for 15.5 hours. After the reaction was completed, the resulted
reaction solution was quenched with a saturated aqueous solution of
ammonium chloride (500 mL), and extracted with dichloromethane (100
mL.times.3). The organic phases were washed with a saturated saline
solution (500 mL), dried with anhydrous sodium sulfate, and
filtered to collect a filtrate. The filtrate was concentrated in
vacuo at 45.degree. C. with a water pump, obtaining a crude
product. The crude product was purified by passing a rapid column
(SiO.sub.2, 100-200 mesh, PE:EA=1:0 to 10:1) to obtain compound
B-1-3. .sup.1H NMR (400 MHz, CHLOROFORM-d) .delta.=7.67 (d, J=2.4
Hz, 1H), 7.35 (dd, J=2.4, 8.4 Hz, 1H), 7.21 (d, J=8.4 Hz, 1H),
6.08-5.91 (m, 1H), 5.39 (q, J=1.6 Hz, 1H), 5.34 (q, J=1.5 Hz, 1H),
5.29-5.24 (q, 1H), 4.57 (s, 2H), 4.13 (td, J=1.3, 5.6 Hz, 2H).
[0105] Step 3: Synthesis of Compound B-1-5.
[0106] Compound B-1-4 (9.9 g, 36.23 mmol, 1 eq) and THF (70.5 mL)
were added to a three-necked flask. After replaced with nitrogen,
the flask was cooled to 0.degree. C., and tert-butyl Grignard
reagent (2M, 29.70 mL, 1.64 eq) was added thereto, and the resulted
mixture was reacted at 0.degree. C. for 1 hour to obtain a first
reaction solution. Compound B-1-3 (12.32 g, 47.09 mmol, 1.3 eq) and
tetrahydrofuran (141 mL) were added to a three-necked flask. After
replaced with nitrogen, the flask was cooled to -78.degree. C., and
N-butyl lithium (2.5M, 21.74 mL, 1.5 eq) was added thereto, and the
resulted mixture was reacted at -78.degree. C. for 0.5 hours to
obtain a second reaction solution. The first reaction solution was
then added dropwise to the second reaction solution using a syringe
to perform reaction at -78.degree. C. for 1 hour and then at
25.degree. C. for 13.5 hours. After the reaction was completed, the
resulted reaction solution was quenched with a saturated aqueous
solution of ammonium chloride (400 mL), then extracted with ethyl
acetate (100 mL.times.3). The organic phases were washed with a
saturated saline solution (1000 mL), dried with anhydrous sodium
sulfate, and filtered to collect a filtrate. The filtrate was
concentrated in vacuo at 45.degree. C. by reducing pressure with a
water pump to obtain a crude product. The crude product was
purified by passing a rapid column (petroleum ether/ethyl acetate
system) to obtain compound B-1-5. .sup.1H NMR (400 MHz,
CHLOROFORM-d) .delta.=8.21 (s, 1H), 7.94 (dd, J=2.0, 8.4 Hz, 1H),
7.48 (d, J=8.2 Hz, 1H), 6.10 (d, J=3.5 Hz, 1H), 6.05-5.94 (m, 1H),
5.38 (dd, J=1.5, 17.2 Hz, 1H), 5.33 (d, J=2.6 Hz, 1H), 5.28-5.23
(m, 1H), 4.65 (s, 2H), 4.63 (br d, J=3.3 Hz, 1H), 4.61 (d, J=3.5
Hz, 1H), 4.15 (d, J=5.5 Hz, 2H), 2.97 (d, J=4.2 Hz, 1H), 1.59 (s,
3H), 1.38 (s, 3H).
[0107] Step 4: Synthesis of Compound B-1-6.
[0108] Compound B-1-5 (8 g, 21.69 mmol, 1 eq), cerium chloride
heptahydrate (9.70 g, 26.03 mmol, 2.47 mL, 1.2 eq) and methanol
(180 mL) were added to a reaction flask. After replaced with
nitrogen, sodium borohydride (1.64 g, 43.38 mmol, 2 eq) was added
thereto at 0.degree. C., and the resulted mixture was reacted at
25.degree. C. for 16 hours. After the reaction was completed, the
reaction solution was quenched with a saturated aqueous solution of
ammonium chloride (250 mL), and a saturated saline solution (250
mL) was added thereto. Then, the reaction solution was extracted
with ethyl acetate (100 mL.times.3) to obtain organic phases (if
the liquid is difficult to separate during extraction, the liquid
can be separated by filtration with diatomite). The organic phases
were dried by anhydrous sodium sulfate and filtered to collect a
filtrate. The filtrate was concentrated to dry at 45.degree. C. by
reducing pressure with a water pump to obtain compound B-1-6.
.sup.1H NMR (400 MHz, CHLOROFORM-d) .delta.=7.61-7.56 (m, 1H),
7.42-7.31 (m, 2H), 6.05-5.92 (m, 2H), 5.41-5.32 (m, 1H), 5.28-5.18
(m, 2H), 4.64-4.59 (m, 2H), 4.49 (d, J=3.5 Hz, 1H), 4.16-4.03 (m,
5H), 3.36 (br s, 1H), 1.46 (s, 3H), 1.30 (s, 3H).
[0109] Step 5: Synthesis of Compound B-1-7.
[0110] Compound B-1-6 (7.2 g, 19.42 mmol, 1 eq), water (45 mL) and
acetic acid (44.31 g, 737.82 mmol, 42.20 mL, 38 eq) were added into
a reaction flask to perform reaction at 100.degree. C. for 7 hours.
After the reaction was completed, the reaction solution was
concentrated to dry at 45.degree. C. by reducing pressure with a
water pump, and then subjected to azeotropic drying with toluene
(100 mL.times.2), obtaining compound B-1-7. .sup.1H NMR (400 MHz,
CHLOROFORM-d) .delta.=7.42 (br d, J=8.6 Hz, 1H), 7.18 (br s, 1H),
7.09 (br d, J=6.8 Hz, 1H), 5.80 (tt, J=6.0, 16.8 Hz, 1H), 5.54-5.08
(m, 4H), 4.58 (br d, J=5.3 Hz, 1H), 4.43 (br s, 2H), 4.08 (br s,
1H), 4.14-3.80 (m, 3H), 3.62-3.28 (m, 3H), 2.20 (br s, 1H).
[0111] Step 6: Synthesis of Compound B-1-8.
[0112] Compound B-1-7 (6 g, 18.14 mmol, 1 eq), triethylamine (12.11
g, 119.72 mmol, 16.66 mL, 6.6 eq) and acetonitrile (110 mL) were
added into a single-necked flask, and then acetic anhydride (12.22
g, 119.72 mmol, 11.21 mL, 6.6 eq) and dimethylaminopyridine (22.16
mg, 181.40 umol, 0.01 eq) were successively added thereto to
perform reaction at 25.degree. C. for 16 hours. After the reaction
was completed, the reaction solution was quenched with a saturated
aqueous solution of sodium bisulfate (100 mL), extracted with ethyl
acetate (50 mL.times.3). The organic phases were washed with a
saturated saline solution (200 mL), dried with anhydrous sodium
sulfate and filtered to collect a filtrate. The filtrate was
concentrated to dry at 45.degree. C. by reducing pressure with a
water pump to obtain a crude product. The crude product was
purified by passing a rapid column (petroleum ether/ethyl acetate
system) to obtain compound B-1-8. .sup.1H NMR (400 MHz,
CHLOROFORM-d) .delta.=7.49 (d, J=1.9 Hz, 1H), 7.33 (d, J=8.3 Hz,
1H), 7.25-7.21 (dd, 1H), 5.99 (tdd, J=5.6, 10.4, 17.2 Hz, 1H), 5.87
(d, J=8.3 Hz, 1H), 5.41-5.36 (m, 1H), 5.36-5.31 (m, 1H), 5.30-5.23
(m, 2H), 5.17-5.10 (t, 1H), 4.61-4.52 (m, 3H), 4.12-4.08 (m, 2H),
2.13-2.10 (s, 3H), 2.07 (s, 3H), 2.04-1.99 (s, 3H), 1.85 (s,
3H).
[0113] Step 7: Synthesis of Compound B-1-9.
[0114] Compound B-1-8 (6.5 g, 13.03 mmol, 1 eq), sodium acetate
(4.28 g, 52.11 mmol, 4 eq), water (13 mL) and glacial acetic acid
(117 mL) were added to a reaction flask. The reaction was cooled to
5.degree. C. after replaced with nitrogen, and palladium dichloride
(5.08 g, 28.66 mmol, 2.2 eq) was added thereto. The resulted
mixture was reacted at 25.degree. C. for 16 hours. After the
reaction was completed, the reaction solution was concentrated to
dry at 45.degree. C. by reducing pressure with a water pump to
obtain a crude product. The crude product was purified by passing a
rapid column (petroleum ether/ethyl acetate system) to obtain
compound B-1-9. .sup.1H NMR (400 MHz, CHLOROFORM-d) .delta.=7.53
(d, J=1.8 Hz, 1H), 7.33 (d, J=8.2 Hz, 1H), 7.21 (dd, J=2.1, 8.3 Hz,
1H), 5.87 (d, J=8.2 Hz, 1H), 5.41-5.34 (t, 1H), 5.30-5.23 (t, 1H),
5.15 (t, J=9.6 Hz, 1H), 4.77 (br d, J=2.4 Hz, 2H), 4.56 (d, J=9.9
Hz, 1H), 2.15-2.10 (s, 3H), 2.07 (s, 3H), 2.02 (s, 3H), 1.85 (s,
3H).
[0115] Step 8: Synthesis of Compound B-1.
[0116] Compound B-1-9 (1 g, 2.18 mmol, 14.04 .mu.L, 1 eq),
triphenylphosphine (857.44 mg, 3.27 mmol, 1.5 eq) and
dichloromethane (20 mL) were added to a reaction flask, and stirred
for half an hour after protected with nitrogen, and then
N-bromosuccinimide (581.85 mg, 3.27 mmol, 1.5 eq) was added thereto
at 0.degree. C. The resulted mixture was reacted at 25.degree. C.
for 15.5 hours. After the reaction was completed, the reaction
solution was concentrated to dry at 25.degree. C. to obtain a crude
product. The crude product was purified by passing a rapid column
(petroleum ether/ethyl acetate system) to obtain compound B-1.
.sup.1H NMR (400 MHz, CHLOROFORM-d) .delta.=7.44-7.36 (m, 2H), 7.29
(s, 1H), 5.87 (d, J=8.2 Hz, 1H), 5.41-5.34 (t, 1H), 5.30-5.23 (m,
1H), 5.15-5.03 (m, 1H), 4.68-4.59 (d, 1H), 4.53 (t, J=9.9 Hz, 2H),
2.22 (s, 1H), 2.13 (s, 2H), 2.08-2.05 (m, 3H), 2.04-2.01 (m, 3H),
1.91-1.86 (m, 3H).
Reference Example 10: Fragment B-2
##STR00032##
[0118] Synthesis Route:
##STR00033## ##STR00034##
[0119] Step 1: Synthesis of Compound B-2-2
[0120] Lithium aluminum hydride (11 g, 289.82 mmol, 1.25 eq) was
dissolved in tetrahydrofuran (200 mL) at 0.degree. C., and filled
with nitrogen protection after replaced with nitrogen three times.
Compound B-2-1 (50 g, 232.51 mmol, 1 eq) was dissolved in
tetrahydrofuran (200 mL), and the resulted solution was slowly
added to the reaction solution at 0.degree. C. Bubbles were
observed and the reaction solution was heated to 25.degree. C. to
continue reaction for 2 hours. Water (11 mL) was slowly added
dropwise thereto at 0.degree. C., then 15% aqueous solution of
sodium hydroxide (11 mL) was added thereto, and finally water (33
mL) was added thereto. The resulted mixture was filtered to obtain
a residue which was then washed twice with ethyl acetate, and the
filtrate was concentrated in vacuo to obtain a crude compound
B-2-2.
[0121] Step 2: Synthesis of Compound B-2-3
[0122] Compound B-2-2 (47.9 g, 238.24 mmol, 1 eq) was dissolved in
dimethylformamide (120 mL), and sodium hydride (14.29 g, 357.36
mmol, 60% purity, 1.5 eq) was added thereto at 0.degree. C.
Stirring was performed at 25.degree. C. for 0.5 hours, and then
3-bromopropene (57.64 g, 476.47 mmol, 41.17 mL, 2 eq) was slowly
added to the reaction solution to continue the reaction at
25.degree. C. for 2 hours. After the reaction was completed, the
reaction solution was quenched with water (50 mL) at 0.degree. C.,
extracted with ethyl acetate (500 mL.times.2), washed with water
(50 mL.times.2) and then washed with a saturated saline solution
(50 mL.times.2), and dried with anhydrous sodium sulfate to obtain
a crude product. The crude product was purified by column
chromatography (petroleum ether/ethyl acetate system) to obtain the
target compound B-2-3.
[0123] Step 3: Synthesis of Compound B-2-4
[0124] Compound B-2-3 (18.5 g, 76.72 mmol, 1.2 eq) was dissolved in
tetrahydrofuran (100 mL) at -78.degree. C., and then n-butyllithium
(2.5 M, 33.25 mL, 1.3 eq) was added under nitrogen protection to
perform reaction at -78.degree. C. for 0.5 hours, obtaining alkyl
lithium solution. Compound B-1-4 (17.47 g, 63.93 mmol, 1 eq) was
dissolved in tetrahydrofuran (100 mL), cooled to 0.degree. C. and
tert-butyl magnesium chloride (1.7 M, 41.37 mL, 1.1 eq) was added
dropwise under nitrogen protection to perform reaction at 0.degree.
C. for 0.5 hours, obtaining a magnesium alkoxy solution. The
magnesium alkoxy solution was slowly added to the alkyl lithium
solution at -78.degree. C. to perform reaction at -78.degree. C.
for 0.5 hours, then the temperature was raised to 25.degree. C. to
continue reaction for 15.5 hours. After the reaction was completed,
an amine chloride solution (50 mL) was added to the resulted
reaction solution at 0.degree. C. and ethyl acetate (200 mL) was
added to dilute the reaction solution. Then the reaction solution
was washed with water (50 mL.times.2) to obtain organic phases. The
combined organic phases were washed with saturated saline solution
(50 mL.times.2) to remove the residue water. Then the resulted was
dried with anhydrous sodium sulfate, filtered and dried by rotation
to obtain a crude product. The crude product was purified by column
chromatography (petroleum ether/ethyl acetate system) to obtain the
target compound B-2-4.
[0125] Step 4: Synthesis of Compound B-2-5
[0126] Compound B-2-4 (17.80 g, 51.09 mmol, 1 eq) was dissolved in
methanol (100 mL), cooled to 0.degree. C., and cerium trichloride
heptahydrate (22.84 g, 61.31 mmol, 5.83 mL, 1.2 eq) and sodium
borohydride (3.87 g, 102.18 mmol, 2 eq) were added thereto
successively, raised to 25.degree. C. to perform reaction for 16
hours. After the reaction was completed, the resulted reaction
solution was quenched with water (30 mL) and concentrated in vacuo.
Then the resulted was diluted with ethyl acetate (100 mL) and
washed with water (50 mL.times.2). Water was removed by using a
saturated saline solution (50 mL.times.2). Finally, the solution
was dried with anhydrous sodium sulfate, filtered and concentrated
to dry by reducing pressure, obtaining the target compound
B-2-5.
[0127] Step 5: Synthesis of Compound B-2-6
[0128] Compound B-2-5 (10.22 g, 29.17 mmol, 1 eq) was dissolved in
water (100 mL) and glacial acetic acid (100 mL) to perform reaction
at 100.degree. C. for 16 hours. After the reaction was completed,
the solvent was vacuum dried by rotation at 60.degree. C., and then
dried with toluene three times to obtain compound B-2-6.
[0129] Step 6: Synthesis of Compound B-2-7
[0130] Compound B-2-6 (9.52 g, 30.68 mmol, 1 eq) and acetic
anhydride (25.05 g, 245.41 mmol, 22.98 mL, 8 eq) were dissolved in
pyridine (40 mL), and stirred at 25.degree. C. for 16 hours. After
the reaction was completed, the reaction solution was diluted with
ethyl acetate (200 mL), washed with 1M dilute hydrochloric acid
(100 mL.times.4). The organic phase was collected and washed with
water (50 mL.times.2), and then washed with a saturated saline
solution (50 mL.times.2), and finally was dried with anhydrous
sodium sulfate, and filtered and concentrated to dry by reducing
pressure, obtaining a crude product. The crude product was purified
by column chromatography (petroleum ether/ethyl acetate system) to
obtain the target compound B-2-7.
[0131] Step 7: Synthesis of Compound B-2-8
[0132] Compound B-2-7 (7 g, 14.63 mmol, 1 eq) and potassium acetate
(5.74 g, 58.52 mmol, 4 eq) were dissolved in acetic acid (135 mL)
and water (15 mL). Palladium dichloride (5.71 g, 32.18 mmol, 2.2
eq) was added in an ice bath under nitrogen protection to perform
reaction at 25.degree. C. for 16 hours. After the reaction was
completed, the reaction solution was vacuum dried by rotation at
45.degree. C. to obtain a crude product. The crude product was
purified by column chromatography (petroleum ether/ethyl acetate
system) to obtain the target compound B-2-8.
[0133] Step 8: Synthesis of Compound B-2
[0134] Compound B-2-8 (2.5 g, 5.70 mmol, 1 eq) was dissolved in
dichloromethane (40 mL), then triphenylphosphine (2.24 g, 8.55
mmol, 1.5 eq) was added thereto, and stirring was performed under
nitrogen protection for 30 minute. The mixture was cooled to
0.degree. C., then N-bromosuccinimide (1.52 g, 8.55 mmol, 1.5 eq)
was added thereto, and stirring was performed at 25.degree. C. for
2.5 hours. After the reaction was completed, the reaction solution
was concentrated to dry at 25.degree. C. to obtain a crude product.
The crude product was purified by column chromatography (petroleum
ether/ethyl acetate system) to obtain the target compound B-2.
.sup.1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 1.85 (s, 3H), 2.01
(s, 3H), 2.1 (s, 3H), 2.19 (s, 3H), 2.37 (s, 3H) 4.43-4.50 (m, 2H),
4.80-4.83 (d, J=10.4 Hz, 1H), 5.055-5.104 (m, 1H), 5.214-5.249 (m,
1H), 5.553-5.602 (m, 1H), 6.444-6.453 (m, 1H), 7.145-7.165 (m, 1H),
7.209-7.224 (m, 1H), 7.251-7.270 (m, 1H).
Reference Example 11: Fragment B-3
##STR00035##
[0136] Synthesis Route:
##STR00036##
[0137] Step 1: Synthesis of Compound B-3-1
[0138] Compound B-2-7 (8.8 g, 18.39 mmol, 1 eq) was dissolved in
1,4-dioxane (100 mL), and thiourea (4.20 g, 55.17 mmol, 3 eq) was
added thereto. After replaced with nitrogen three times,
trimethylsilyl trifluoromethanesulfonate (14.31 g, 64.37 mmol, 3.5
eq) was added thereto at 25.degree. C., and the resulted was heated
to 60.degree. C. to perform reaction for 2 hours, and then cooled
to 25.degree. C. Methyl iodide (13.30 g, 93.70 g mmol, 5.09 eq) and
diisopropylethylamine (19.02 g, 147.13 mmol, 8 eq) were added
successively thereto to perform reaction at 25.degree. C. for 14
hours. After the reaction was completed, the reaction solution was
diluted with water (80MI), extracted with ethyl acetate (80
mL.times.3) to collect organic phases. The combined organic phases
were with a saturated saline solution (50 mL), dried with anhydrous
sodium sulfate, and filtered to obtain a filtrate. The filtrate was
dried by rotation under reduced pressure to obtain a crude product.
The crude product was purified by column chromatography (petroleum
ether/ethyl acetate system) to obtain the target compound B-3-1,
which was confirmed by LCMS.
[0139] Step 2: Synthesis of Compound B-3-2
[0140] Compound B-3-1 (2 g, 4.29 mmol, 1 eq), barbituric acid (1.10
g, 8.57 mmol, 2 eq) and ethanol (20 mL) were added to a reaction
flask. After replaced with nitrogen three times,
tetra-triphenylphosphine palladium (495.37 mg, 428.68 .mu.mol, 0.1
eq) was added thereto to perform reaction at 70.degree. C. under
nitrogen atmosphere for 16 hours. After the reaction was completed,
the reaction solution was diluted with water (20 mL) and extracted
with ethyl acetate (20 mL.times.3). The combined organic phases
were washed with a saturated saline solution (20 mL), dried with
anhydrous sodium sulfate, and filtered to obtain a filtrate. The
filtrate was dried by rotation under reduced pressure to obtain a
crude product. The crude product was purified by column
chromatography (petroleum ether/ethyl acetate system) to obtain the
target compound B-3-2, which was confirmed by LCMS.
[0141] Step 3: Synthesis of Compound B-3
[0142] Compound B-3-2 (1.5 g, 3.52 mmol, 1 eq), triphenylphosphine
(1.38 g, 5.28 mmol, 1.5 eq) and dichloromethane (20 mL) were added
to a reaction flask. After replacing nitrogen three times, stirring
was performed at 25.degree. C. for 0.5 hours, then
N-bromosuccinimide (938.98 mg, 5.28 mmol, 1.5 eq) was added thereto
at 0.degree. C. to perform reaction at 25.degree. C. for 1.5 hours.
After the reaction was completed, the reaction solution was diluted
with water (20 mL) and extracted with ethyl acetate (20
mL.times.3). The combined organic phases were dried with anhydrous
sodium sulfate, filtered to obtain a filtrate. The filtrate was
dried by rotation under reduced pressure to obtain a crude product.
The crude product was purified by column chromatography (petroleum
ether/ethyl acetate system) to obtain the target compound B-3.
.sup.1H NMR (400 MHz, CHLOROFORM-d) .delta.=7.25 (d, J=6.4 Hz, 2H),
7.18 (d, J=8.4 Hz, 1H), 5.38 (t, J=9.6 Hz, 1H), 5.25 (t, J=9.6 Hz,
1H), 5.13 (t, J=9.6 Hz, 1H), 4.56 (d, J=9.6 Hz, 1H), 4.53 (q, 10.4
Hz, 2H), 4.43 (d, J=9.6 Hz, 1H), 2.40 (s, 3H), 2.21 (s, 3H), 2.11
(s, 3H), 2.02 (s, 3H), 1.84 (s, 3H).
[0143] The fragment B-4 in the following table is synthesized with
reference to the steps 1-8 in Reference Example 9. The fragment B-5
in the following table is synthesized with reference to the steps
1-8 in Reference Example 10. The structures in the table also
include their possible isomers.
TABLE-US-00002 Reference Fragments Example B Structure .sup.1H NMR
12 B-4 ##STR00037## .sup.1H NMR (400 M Hz, CHLOROFORM-d) .delta. =
7.33-7.27 (m, 2H), 6.85 (m, J = 8.4 Hz, 1H), 5.87 (d, J = 8.2 Hz,
1H), 5.40-5.32 (t, 1H), 5.31-5.22 (m, 1H), 5.15 (m, J = 9.7 Hz,
1H), 4.64-4.54 (m, 1H), 4.48 (dd, J = 9.8, 13.6 Hz, 2H), 3.92-3.85
(d, 3H), 2.21 (s, 1H), 2.11 (s, 2H), 2.08-2.04 (d, 3H), 2.04-1.99
(d, 3H), 1.89- 1.83 (d, 3H) 13 B-5 ##STR00038## .sup.1H NMR (400 M
Hz, CHLOROFORM-d) .delta. ppm 1.85 (s, 1.5H) 1.87 (s, 1.5H) 2.00
(s, 1.5H) 2.02 (s, 1.5H) 2.04 (s, 1.5H) 2.06 (s, 1.5H) 2.09-2.13
(m, 1.5H) 2.20 (s, 1.5H) 4.39-4.46 (m, 1H) 4.49 (br s, 0.5H) 4.53
(d, J = 10.29 Hz, 1H) 4.84 (d, J = 10.04 Hz, 0.5H) 5.01-5.14 (m,
1H) 5.20-5.29 (m, 1H) 5.33-5.39 (m, 0.5H) 5.58 (t, J = 9.91 Hz,
0.5H) 5.86 (d, J = 8.28 Hz, 0.5H) 6.45 (d, J = 3.76 Hz, 0.5H) 7.04
(td, J = 9.03, 2.26 Hz, 1H) 7.29-7.32 (m, 1H) 7.34-7.37 (m, 1H)
[0144] The fragment B-6 in the following table is synthesized with
reference to the steps 1-3 in Reference Example 11. The structures
in the table also include their possible isomers.
TABLE-US-00003 Reference Example Fragment Structure NMR 14 B-6
##STR00039## .sup.1H NMR (400 M Hz, CHLOROFORM-d) .delta. ppm
7.28-7.32 (m, 1H) 7.26 (s, 1H) 7.18-7.25 (m, 1H) 5.31-5.41 (m, 1H)
5.19-5.26 (m, 1H) 5.12 (t, J = 9.69 Hz, 1H) 4.47-4.60 (m, 3H) 4.43
(d, J = 9.88 Hz, 1H) 2.76 (q, J = 7.63 Hz, 2H) 2.17-2.25 (m, 3H)
2.07-2.14 (m, 3H) 1.98-2.06 (m, 3H) 1.80-1.90 (m, 3H) 1.28 (t, J =
7.57 Hz, 3H)
Example 1: WXD001
##STR00040##
[0146] Synthesis Route:
##STR00041##
[0147] Step 1: Synthesis of Compound WXD001-1.
[0148] Compound B-1 (1 g, 1.92 mmol, 1 eq) was mixed with compound
A-1 (797.78 mg, 2.49 mmol, 1 mL, 1.3 eq), sodium carbonate (2 M,
1.92 mL, 2 eq), toluene (20 mL), ethanol (5 mL) and water (5 mL).
After purging nitrogen, tetra-triphenylphosphine palladium (221.48
mg, 191.67 .mu.mol, 0.1 eq) was added thereto to perform reaction
at 50.degree. C. for 16 hours, and the reaction solution turned
black. After the reaction was completed, the reaction solution was
concentrated with a water pump under reduced pressure at 45.degree.
C. to remove ethanol, and then concentrated with an oil pump to
remove toluene and water, obtaining a crude product. The crude
product was purified by column chromatography (petroleum
ether/ethyl acetate system) to obtain the target compound WXD001-1.
.sup.1H NMR (400 MHz, CHLOROFORM-d) .delta.=7.37 (d, J=8.2 Hz, 1H),
7.30 (d, J=8.2 Hz, 2H), 7.20 (dd, J=2.1, 8.3 Hz, 1H), 7.16-7.07 (m,
3H), 5.89 (dd, J=2.1, 3.4 Hz, 1H), 5.84 (d, J=8.2 Hz, 1H),
5.38-5.29 (m, 1H), 5.25 (dd, J=8.3, 9.6 Hz, 1H), 5.09 (t, J=9.6 Hz,
1H), 4.47 (d, J=9.7 Hz, 1H), 4.12-4.00 (m, 2H), 2.76-2.65 (m, 4H),
2.21-2.14 (m, 2H), 2.13-2.08 (m, 3H), 2.07-2.05 (m, 3H), 2.03-2.00
(m, 3H), 1.74-1.69 (m, 3H).
[0149] Step 2: Synthesis of Compound WXD001-2.
[0150] Compound WXD001-1 (1 g, 1.57 mmol, 1 eq), thiourea (239.73
mg, 3.15 mmol, 2 eq) and dioxane (12 mL) were added to a reaction
flask. After purging nitrogen, trimethylsilyl
trifluoromethanesulfonate (874.97 mg, 3.94 mmol, 711.35 uL, 2.5 eq)
was added thereto, and slowly heated to 80.degree. C. to perform
reaction for 2 hours. After cooling to 25.degree. C.,
diisopropylethylamine (1.02 g, 7.87 mmol, 1.37 mL, 5 eq) and methyl
iodide (670.52 mg, 4.72 mmol, 294.09 uL, 3 eq) were added
successively thereto to perform reaction 25.degree. C. for 14
hours. After the reaction was completed, the reaction solution was
diluted with water (5 mL), extracted with dichloromethane (2
mL.times.3) to obtain organic phases. The organic phases were
washed with a saturated saline solution (10 mL), dried with
anhydrous sodium sulfate, and filtered to obtain a filtrate. The
filtrate was concentrated to dry at 45.degree. C. with a water pump
to obtain a crude product. The crude product was purified by column
chromatography (petroleum ether/ethyl acetate system) to obtain the
target compound WXD001-2. .sup.1H NMR (400 MHz, CHLOROFORM-d)
.delta.=7.37 (d, J=8.2 Hz, 1H), 7.30 (d, J=8.2 Hz, 2H), 7.19 (dd,
J=2.0, 8.3 Hz, 1H), 7.14-7.08 (m, 3H), 5.89 (br s, 1H), 5.31 (s,
1H), 5.19 (s, 1H), 5.04 (s, 1H), 4.50 (d, J=9.9 Hz, 1H), 4.38 (d,
J=9.9 Hz, 1H), 4.08 (d, J=17.0 Hz, 2H), 2.69 (m, J=6.0, 8.1 Hz,
4H), 2.24-2.16 (m, 2H), 2.15 (s, 3H), 2.09 (s, 3H), 2.00 (s, 3H),
1.71 (s, 3H).
[0151] Step 3: Synthesis of Compound WXD001.
[0152] Compound WXD001-2 (760 mg, 1.22 mmol, 1 eq), methanol (6 mL)
and tetrahydrofuran (3 mL) were added to a reaction flask, and then
lithium hydroxide monohydrate (1.02 g, 24.39 mmol, 20 eq) and water
(6 mL) were added thereto to perform reaction at 25.degree. C. for
16 hours. After the reaction was completed, the reaction solution
was diluted with water (10 mL), extracted with ethyl acetate (10
mL.times.3) to obtain organic phases. The organic phases were
washed with a saturated saline solution (30 mL), dried with
anhydrous sodium sulfate, and filtered to obtain a filtrate. The
filtrate was concentrated to dry at 45.degree. C. with a water pump
to obtain a crude product. The crude product, was purified by
preparative high performance liquid chromatography
(acetonitrile/water-aqua ammonia system) to obtain the target
compound WXD001. SFC showed that the enantiomeric excess ratio was
100%. .sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta.=7.37 (d,
J=8.2 Hz, 1H), 7.34-7.30 (m, 2H), 7.28-7.23 (m, 2H), 7.16 (d, J=8.4
Hz, 2H), 6.00-5.84 (m, 1H), 4.38 (d, J=9.5 Hz, 1H), 4.14 (d, J=9.5
Hz, 1H), 4.11-4.04 (d, 2H), 3.48-3.42 (t, 1H), 3.39-3.32 (m, 2H),
2.72-2.63 (m, 4H), 2.23-2.12 (m, 2H), 2.12 (s, 3H).
Example 2: WXD002
##STR00042##
[0154] Synthesis Route:
[0155] Step 1: Synthesis of Compound WXD002-1.
[0156] Compound B-3 (40 mg, 81.74 .mu.mol, 1 eq), compound A-3
(41.96 mg, 122.61 .mu.mol, 1.5 eq), sodium carbonate (17.33 mg,
163.47 .mu.mol, 2 eq), toluene (3 mL), ethanol (0.3 mL) and water
(0.3 mL) were added to a reaction flask. After purging nitrogen
three times, tetra-triphenylphosphine palladium (9.45 mg, 8.17
.mu.mol, 0.1 eq) was added thereto to perform reaction at
50.degree. C. for 5 hours under a nitrogen atmosphere. After the
reaction was completed, the reaction solution was diluted with
water (5 mL), extracted with ethyl acetate (5 mL.times.3) to obtain
organic phases. The combined organic phases were dried with
anhydrous sodium sulfate, filtered to obtain a filtrate. The
filtrate was dried by rotation under reduced pressure to obtain a
crude product. The crude product was purified by preparative TLC
(petroleum ether/ethyl acetate system) to obtain the target
compound WXD002-1. .sup.1H NMR (400 MHz, CHLOROFORM-d) .delta.=7.30
(d, J=8.0 Hz, 2H), 7.17-7.12 (m, 2H), 7.03-6.99 (m, 3H), 5.96 (t,
J=3.6 Hz, 1H), 5.35 (t, J=9.2 Hz, 1H), 5.23 (t, J=9.6 Hz, 1H), 5.14
(t, J=9.6 Hz, 1H), 4.53 (d, 10.0 Hz, 1H), 4.39 (d, 10.0 Hz, 1H),
4.02 (s, 4H), 3.94 (d, J=6.8 Hz, 2H), 2.66-2.62 (m, 2H), 2.46-2.45
(m, 2H), 2.02 (s, 3H), 2.17 (s, 3H), 2.10 (s, 3H), 2.01 (s, 3H),
1.93 (t, J=6.4 Hz, 2H), 1.74 (s, 3H).
[0157] Step 2: Synthesis of WXD002
[0158] Compound WXD002-1 (42 mg, 67.23 .mu.mol, 1 eq), methanol (1
mL), tetrahydrofuran (0.5 mL), water (1 mL), and lithium hydroxide
monohydrate (56.42 mg, 1.34 mmol, 20 eq) were added to a reaction
flask to perform reaction at 25.degree. C. for 1 hour. After the
reaction was completed, the reaction solution was diluted with
water (5 mL) and extracted with ethyl acetate (5 mL.times.4) to
obtain organic phases. The combined organic phases were dried over
anhydrous sodium sulfate, and filtered to obtain a filtrate. The
filtrate was dried by rotation under reduced pressure to obtain a
crude product. The crude product was purified by preparative high
performance liquid chromatography mechanical separation
(acetonitrile/water-aqua ammonia system) to obtain the target
compound WXD002. .sup.1H NMR (400 MHz, CHLOROFORM-d) .delta.=7.30
(d, J=8.4, 2H), 7.18-7.13 (m, 3H), 7.09 (d, J=8.0 Hz, 2H), 5.95 (t,
J=3.6 Hz, 1H), 4.40 (d, J=9.6 Hz, 1H), 4.14 (d, J=9.2 Hz, 1H),
3.99-3.97 (m, 6H), 3.48-3.35 (m, 3H), 2.62-2.58 (m, 2H), 2.41 (s,
2H), 2.21 (s, 3H), 2.13 (s, 3H), 1.89 (t, J=6.4 Hz, 2H).
[0159] Each fragment of the Examples 3-9 in the following table 1
was synthesized with reference to the steps 1-3 in Example 1. The
structures in table 1 also include their possible isomers.
TABLE-US-00004 TABLE 1 Reference Example Fragment A Fragment B
Compound Structure 3 ##STR00043## ##STR00044## WXD003 ##STR00045##
4 ##STR00046## ##STR00047## WXD004 ##STR00048## 5 ##STR00049##
##STR00050## WXD005 ##STR00051## 6 ##STR00052## ##STR00053## WXD006
##STR00054## 7 ##STR00055## ##STR00056## WXD007 ##STR00057## 8
##STR00058## ##STR00059## WXD008 ##STR00060## 9 ##STR00061##
##STR00062## WXD009 ##STR00063##
[0160] Each fragment in Example 10 in the following table 2 was
synthesized with reference to the steps 1-2 in Example 2. The
structures in table 2 also include their possible isomers.
TABLE-US-00005 TABLE 2 Reference Example Fragment A Fragment B
Compound Structure 10 ##STR00064## ##STR00065## WXD010
##STR00066##
[0161] The hydrogen spectrum and mass spectrum data of each example
are shown in Table 3.
TABLE-US-00006 TABLE 3 Reference Example Compound NMR MS m/z 1
WXD001 .sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta. = 7.37 (d, J
= 8.2 Hz, 1H), 519 7.34-7.30 (m, 2H), 7.28-7.23 (m, 2H), 7.16 (d, J
= 8.4 Hz, 2H), 6.00-5.84 (M + Na) (m, 1H), 4.38 (d, J = 9.5 Hz,
1H), 4.14 (d, J = 9.5 Hz, 1H), 4.11-4.04 (d, 2H), 3.48-3.42 (t,
1H), 3.39-3.32 (m, 2H), 2.72-2.63 (m, 4H), 2.23- 2.12 (m, 2H), 2.12
(s, 3H). 2 WXD002 .sup.1H NMR (400 MHz, CHLOROFORM-d) .delta. =
7.30 (d, J = 8.4, 2H), 521 7.18-7.13 (m, 3H), 7.09 (d, J = 8.0 Hz,
2H), 5.95 (t, J = 3.6 Hz, 1H), (M + Na) 4.40 (d, J = 9.6 Hz, 1H),
4.14 (d, J = 9.2 Hz, 1H), 3.99-3.97 (m, 6H), 3.48-3.35 (m, 3H),
2.62-2.58 (m, 2H), 2.41 (s, 2H), 2.21 (s, 3H), 2.13 (s, 3H), 1.89
(t, J = 6.4 Hz, 2H). 3 WXD003 .sup.1H NMR (400 MHz,
METHANOL-d.sub.4) .delta. ppm 2.07-2.18 (m, 5 H), 499 2.20 (s, 3
H), 2.61-2.74 (m, 4 H), 3.33-3.49 (m, 3 H), 3.98 (s, 2 H), (M + Na)
4.12 (d, J = 9.03 Hz, 1 H), 4.38 (d, J = 9.54 Hz, 1 H), 5.90 (br s,
1 H), 7.06-7.20 (m, 5 H), 7.30 (d, J = 8.28 Hz, 2 H). 4 WXD004
.sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta. ppm 2.14 (s, 3 H),
2.21 (s, 465 3 H), 2.49 (br dd, J = 4.44, 2.69 Hz, 2 H), 3.35-3.49
(m, 3 H), 3.91 (t, J = (M + Na) 5.50 Hz, 2 H), 3.99 (s, 2 H), 4.13
(d, J = 9.01 Hz, 1 H), 4.28 (q, J = 2.79 Hz, 2 H), 4.39 (d, J =
9.38 Hz, 1 H), 6.13 (br s, 1 H), 7.11 (d, J = 8.13 Hz, 2 H),
7.14-7.20 (m, 3 H), 7.33 (d, J = 8.25 Hz, 2 H). 5 WXD005 .sup.1H
NMR (400 MHz, METHANOL-d.sub.4) .delta. ppm 1.60-1.70 (m, 2 H), 463
1.74-1.82 (m, 2 H), 2.13 (s, 3 H), 2.15-2.20 (m, 2 H), 2.21 (s, 3
H, 2.32-2.42 (M + Na) (m, 2 H), 3.33-3.51 (m, 3 H), 3.96 (s, 2 H),
4.13 (d, J = 9.03 Hz, 1 H), 4.39 (d, J = 9.54 Hz, 1 H), 6.06 (dt, J
= 3.58, 2.10 Hz, 1 H), 7.06 (d, J = 8.28 Hz, 2 H), 7.11-7.20 (m, 3
H), 7.26 (d, J = 8.28 Hz, 2 H). 6 WXD006 .sup.1H NMR (400 MHz,
CHLOROFORM-d) .delta. ppm 7.09-7.17 (m, 3 H), 505 6.68 (d, J = 3.5
Hz, 1 H), 6.50 (d, J = 3.5 Hz, 1 H), 5.77 (br s, 1 H), (M + Na)
4.33 (br d, J = 9.5 Hz, 1 H), 4.13 (br d, J = 9.0 Hz, 1 H), 4.01
(s, 2 H), 3.61-3.70 (m, 1 H), 3.45-3.56 (m, 2 H), 2.78 (br s, 1 H),
2.41-2.66 (m, 5 H), 2.22 (s, 3 H), 2.13 (s, 3 H), 1.99-2.11 (m, 2
H), 1.95 (br s, 1 H) 7 WXD007 .sup.1H NMR (400 MHz,
METHANOL-d.sub.4) .delta. ppm 2.15 (s, 3 H), 2.28 471 (s, 3 H),
2.45 (br d, J = 1.51 Hz, 2 H), 3.35-3.50 (m, 3 H), 3.86 (t, J = (M
+ Na) 5.52 Hz, 2 H), 4.10 (s, 2 H), 4.14 (d, J = 9.29 Hz, 1 H),
4.21 (br d, J = 2.51 Hz, 2 H), 4.39 (d, J = 9.54 Hz, 1 H), 5.97 (br
s, 1 H), 6.61 (d, J = 3.51 Hz, 1 H), 6.81 (d, J = 3.76 Hz, 1 H),
7.13-7.17 (m, 1 H), 7.18-7.21 (m, 1 H), 7.23 (s, 1 H). 8 WXD008
.sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta. = 7.28 (d, J = 8.2
Hz, 2H), 515 7.23 (dd, J = 2.1, 8.4 Hz, 1H), 7.19-7.12 (m, 3H),
6.93 (d, J = 8.4 Hz, (M + Na) 1H), 5.90 (br s, 1H), 4.37 (d, J =
9.4 Hz, 1H), 4.08 (d, J = 9.0 Hz, 1H), 3.97-3.89 (m, 2H), 3.81 (s,
3H), 3.50-3.36 (m, 3H), 2.71-2.62 (m, 4H), 2.20-2.13 (m, 2H), 2.12
(s, 3H) 9 WXD009 .sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta.
ppm 2.06 (s, 3 H), 2.07-2.09 503 (m, 2 H), 2.54-2.69 (m, 4 H),
3.29-3.45 (m, 3 H), 3.94 (d, J = 2.76 Hz, 2 H), (M + Na) 4.09 (d, J
= 9.54 Hz, 1 H), 4.34 (d, J = 9.54 Hz, 1 H), 5.87 (br s, 1 H),
6.96-7.04 (m, 1 H), 7.14 (d, J = 8.28 Hz, 2 H), 7.18-7.25 (m, 2 H),
7.28 (d, J = 8.28 Hz, 2 H). 10 WXD010 .sup.1H NMR (400 MHz,
METHANOL-d.sub.4) .delta. ppm 1.09 (t, J = 7.53 Hz, 3 H), 513 2.10
(m, 1 H), 2.13 (s, 3 H), 2.14-2.22 (m, 1 H), 2.56-2.74 (m, 6 H),
3.34-3.48 (M + Na) (m, 3 H), 4.02 (s, 2 H), 4.13 (d, J = 9.03 Hz, 1
H), 4.39 (d, J = 9.29 Hz, 1 H), 5.91 (br s, 1 H), 7.10 (d, J = 8.28
Hz, 2 H), 7.14-7.21 (m, 2 H), 7.21- 7.25 (m, 1 H), 7.30 (d, J =
8.28 Hz, 2 H).
Experiment Example 1. In Vitro Cell Viability Test
[0162] Experimental Steps and Methods:
[0163] Biological Activity Experiment 1: SGLT1 Glucose Transport
Test.
[0164] 1. Purpose of the Experiment:
[0165] The effect of the compound on the glucose transport activity
of SGLT1 transporter was detected by measuring the amount of
['C]-labeled glucose entering cells with high expression of
Human-SGLT1.
[0166] 2. Experimental Methods.
[0167] 2.1. Cell Preparation.
[0168] The cells stably expressing Human-SGLT1 were constructed by
Wuxi AppTec(Shanghai) Co., Ltd. SGLT1 cells were laid in Cytostar-T
(PerkinElmer) 96-well cell culture plate and cultured overnight in
5% CO.sub.2 environment at 37.degree. C.
[0169] 2.2. SGLT1 Glucose Transport Test.
[0170] 1) Experimental buffer, including: 10 mM
4-hydroxyethylpiperazineethanesulfonic acid (HEPES), 1.2 mM
magnesium chloride (MgCl.sub.2), 4.7 mM potassium chloride (KCl),
2.2 mM calcium chloride (CaCl.sub.2) and 120 mM sodium chloride
(NaCl).
[0171] 2) Compounds were serially diluted in 100% dimethyl
sulfoxide (DMSO) starting at 1 mM and make a 5-folds, 8 points
serials compound dilutions.
[0172] 3) 3 .mu.M ['.sup.4C]-labeled methyl a-D-glucopyranoside was
prepared with the experimental buffer.
[0173] 4) The cells were treated with 49 uL of the experimental
buffer, 1 .mu.L of the gradient diluted compound and 50 .mu.L of
the 3 .mu.M [.sup.14C]-labeled methyl a-D-lucopyranosid solution at
37.degree. C. for 2 hours.
[0174] 5) The data were read with an isotope detector Micro beta
Reader.
[0175] 6) The IC.sub.50 value of the tested compound was obtained
by the calculation formula: log(inhibitor) vs. response--Variable
slope, using GraphPad Prism 5.0 software.
[0176] Biological Activity Experiment 2: SGLT2 Glucose Transport
Test.
[0177] 1. Purpose of the Experiment:
[0178] The effect of the compound on the glucose transport activity
of SGLT2 transporter was detected by measuring the amount of
[.sup.14C]-labeled glucose entering cells with high expression of
Human-SGLT2.
[0179] 2. Experimental Methods.
[0180] 2.1. Cell Preparation.
[0181] The cells stably expressing Human-SGLT2 were constructed by
Wuxi AppTec(Shanghai) Co., Ltd. SGLT2 cells were laid in 96-well
cell culture plate (Greiner) and cultured overnight in 5% CO.sub.2
environment at 37.degree. C.
[0182] 2.2. SGLT2 Glucose Transport Test.
[0183] 1) Experimental buffer, including: 10 mM
4-hydroxyethylpiperazineethanesulfonic acid (HEPES), 1.2 mM
magnesium chloride (MgCl.sub.2), 4.7 mM potassium chloride (KCl),
2.2 mM calcium chloride (CaCl.sub.2) and 120 mM sodium chloride
(NaCl).
[0184] 2) Stop buffer, including: 10 mM
4-hydroxyethylpiperazineethanesulfonic acid (HEPES), 1.2 mM
magnesium chloride (MgCl.sub.2), 4.7 mM potassium chloride (KCl),
2.2 mM calcium chloride (CaCl.sub.2), 120 mM sodium chloride (NaCl)
and 1 .mu.M LX4211.
[0185] 3) Compounds were serially diluted in 100% dimethyl
sulfoxide (DMSO) starting at 10 uM and make a 5-folds, 8 points
serials compound dilutions.
[0186] 4) 6 .mu.M [.sup.14C]-labeled methyl a-D-lucopyranosid was
prepared with the experimental buffer.
[0187] 5) The cells were treated with 49 uL of the experimental
buffer, 1 .mu.L of the gradient diluted compound and 50 .mu.L of
the 6 .mu.M [.sup.14C]-labeled methyl a-D-lucopyranosid solution at
37.degree. C. for 2 hours.
[0188] 6) The solution was sucked out from holes, and the cells
were rinsed with the stop buffer for 3 times.
[0189] 7) The cells were lysed with 50 ul of 10% sodium hydroxide
solution, the cell lysate was sucked into a scintillation tube,
into which 2 mL scintillation solution was then added.
[0190] 8) The data were read with an isotope detector Tricarb.
[0191] 9) The IC.sub.50 value of the tested compound was obtained
by the calculation formula: log(inhibitor) vs. response--Variable
slope, using GraphPad Prism 5.0 software.
[0192] The experimental results are shown in Table 4:
TABLE-US-00007 TABLE 4 Results of cell viability test in vitro
Human-SGLT1 Human-SGLT2 Compound IC.sub.50 (nM) IC.sub.50 (nM)
Sotagliflozin 69.0 1.15 WXD001 210 3.98 WXD002 55.6 1.01 WXD003
49.5 2.04 WXD004 14.1 1.49 WXD005 21.9 1.78 WXD006 39.2 1.02 WXD007
30.1 1.35 WXD008 17.2 5.40 WXD009 233 4.68 WXD010 14.8 2.49
Conclusion: The Compound of the Present Disclosure Exhibits
Superior In Vitro Inhibitory Activity Against Human-SGLT1 and
Human-SGLT2
Experimental Example 2. Study on the In Vivo Pharmacokinetics in
Animals
[0193] Study on the In Vivo Pharmacokinetics in Rats
[0194] The purpose of the experiment: Male SD rats were used as
test animals and were given a single administration to determine
the plasma concentration of the compound and evaluate the
pharmacokinetic behavior.
[0195] Experimental method: Six healthy adult male SD rats were
divided into 2 groups, with 3 rats in an intravenous injection
group and 3 rats in an oral administration group. In the
intravenous injection group, the test compound was mixed with an
appropriate amount of vehicle (10% N-methyl pyrrolidone (NMP)/10%
polyethylene glycol-15 hydroxystearate (available from solutol)/80%
water), then vortexed and sonicated to prepare 0.2 mg/mL clear
solution, which was filtered by microporous membrane. In the oral
administration group, 10% N-methyl pyrrolidone (NMP)/10%
polyethylene glycol-15 hydroxy stearate (available from
solutol)/80% water was used as a vehicle and mixed with the test
compound, then vortexed and sonicated to prepare 0.40 mg/mL clear
solution. The rats were given intravenous administration at a dose
of 1 mg/kg or oral administration at a dose of 2 mg/kg. Whole blood
was collected at a certain period of time to prepare the plasma.
The drug concentration of the plasma was analyzed by LC-MS/MS
method, and the pharmacokinetic parameters were calculated using
Phoenix WinNonlin software (Pharsight Company, USA).
[0196] The experimental results are shown in Table 5:
TABLE-US-00008 TABLE 5 Results of pharmacokinetic (PK) test of
compounds Oral DNAUC C.sub.max (nM h/ Vd.sub.ss Cl T.sub.1/2
Compound (nM) F % mpk) (L/kg) (mL/min/kg) (h) Sotagliflozin 364
54.8 969 2.01 22.6 1.27 WXD001 410 59.0 1823 3.86 10.1 4.59 WXD003
391 47.7 1013 2.94 14.5 2.56 WXD010 356 56.4 2320 2.48 8.06 4.50
Notes: C.sub.max represents the maximum concentration; F %
represents oral bioavailability; Oral DNAUC = AUC.sub.PO/Dose (unit
oral exposure), AUC.sub.PO represents oral exposure, Dose is drug
dose; Vd.sub.ss is distribution volume; Cl is clearance rate; and
T.sub.1/2 is half-life.
Study on the In Vivo Pharmacokinetics in Beagle Dogs
[0197] The purpose of the experiment: Male beagle (Beagle) dogs
were used as test animals and were given a single administration to
determine the plasma concentration of the compound and evaluate the
pharmacokinetic behavior.
[0198] Experimental method: Six male Beagle dogs were divided into
2 groups, with 3 dogs in an intravenous injection group and 3 dogs
in an oral administration group. In the intravenous injection
group, the test compound was mixed with an appropriate amount of
vehicle (20% polyethylene glycol-400 (PEG400)/10% polyethylene
glycol-15 hydroxy stearate (available from solutol)/70% water),
then vortexed and sonicated to prepare 1 mg/mL clear solution,
which was filtered by microporous membrane. In the oral
administration group, 20% polyethylene glycol-400 (PEG400)/10%
polyethylene glycol-15 hydroxy stearate (available from
solutol)/70% water was used as a vehicle and mixed with the test
compound, then vortexed and sonicated to prepare 1 mg/mL clear
solution. The dogs were given intravenous administration at a dose
of 1 mg/kg or oral administration at a dose of 2 mg/kg. Whole blood
was collected at a certain period of time to prepare the plasma.
The drug concentration of the plasma was analyzed by LC-MS/MS
method, and the pharmacokinetic parameters were calculated using
Phoenix WinNonlin software (Pharsight Company, USA).
[0199] The experimental results are shown in Table 6:
TABLE-US-00009 TABLE 6 Results of pharmacokinetic (PK) test of
compounds Oral DNAUC C.sub.max (nM h/ Vd.sub.ss Cl T.sub.1/2
Compound (nM) F % mpk) (L/kg) (mL/min/kg) (h) WXD010 1287 82 4051
1.49 6.9 2.4 Notes: C.sub.max represents the maximum concentration;
F % represents oral bioavailability; Oral DNAUC = AUC.sub.PO/Dose
(unit oral exposure), AUC.sub.PO represents oral exposure, Dose is
drug dose; Vd.sub.ss is distribution volume; Cl is clearance rate;
T.sub.1/2 is half-life.
Conclusion: The Compound of the Present Disclosure has Good Oral
Exposure and Bioavailability
Experimental Example 3. Study on In Vivo Efficacy of Oral Glucose
Tolerance Test (OGTT) in Rats: Study on the In Vivo Efficacy of
Oral Glucose Tolerance Test (OGTT) in Rats for the First Time
[0200] Summary of the Experiment:
[0201] 1. Animals:
TABLE-US-00010 Animal: Species SD rats Gender: Male Age/weight:
About 8 weeks old/250 g Supplier: Shanghai SLAC Animal feed
Ordinary feed for rats and mice
[0202] 2. Experimental Grouping:
TABLE-US-00011 Number of Administration Administration animals
Groups Compound Dose Frequency method per group 1 Vehicle control
group 0 Single Intragastric 5 administration administration 2
Positive compound 10 mg/kg Single Intragastric 5 (Sotagliflozin)
administration administration 3 WXD001 10 mg/kg Single Intragastric
5 administration administration 4 WXD003 10 mg/kg Single
Intragastric 5 administration administration
[0203] Experiment Procedure:
[0204] 1. Animal Adaptation and Preparation:
[0205] The experimental animals were allowed to adapt to the
environment in the animal room for one week after they arrived.
[0206] 2. Fasting and Drug Administration.
[0207] The animals fasted in the metabolic cage for 18 hours, then
were given drugs or vehicle (2 ml/kg) according to the above table,
and subsequently were immediately given 50% glucose solution (2
g/kg, 4 ml/kg).
[0208] 3. Urine glucose and blood glucose test.
[0209] 2 hours after the administration of glucose solution, feed
intake was restored. Blood samples were taken at 0 min, 15 min, 30
min, 45 min, 60 min and 120 min respectively, and urine samples
were collected at 0-24 h to test blood glucose, urine glucose
(mg/200 g) and urine volume respectively.
[0210] 4. Data Analysis:
[0211] All values are represented as averages. Statistical analysis
was performed using Graphpad Prism 6 one-way analysis of variance
and Tukey's multiple comparison test. P value of less than 0.05 is
considered statistically significant.
[0212] The Experimental Results are Shown in Table 7:
TABLE-US-00012 TABLE 7 Results of glucose tolerance test in rats.
Vehicle Positive control compound Compound group (Sotagliflozin)
WXD001 WXD003 OGTT blood 1033 823* 832* 797** glucose level
AUC.sub.0-2 hr (mol/L .times. min) Urine glucose 1.8 2424.0****
2181.4**** 1636.4**** level (mg/200 g, BW) Urine volume 21.36
39.74**** 37.58**** 24.58 (mL/200 gBW) Notes: *p < 0.05, **p
< 0.01, **p < 0.001, ****p < 0.0001 vs. vehicle control
group.
[0213] Study on the In Vivo Efficacy of Oral Glucose Tolerance Test
(OGTT) in Rats for the Second Time:
[0214] Summary of the Experiment:
[0215] 1. Animals:
TABLE-US-00013 Animal: Species SD rats Gender: Male Age/weight:
About 8 weeks old/250 g Supplier: Shanghai SLAC Animal feed
Ordinary feed for rats and mice
[0216] 2. Experiment Grouping:
TABLE-US-00014 Number of Administration Administration animals
Groups Compound Dose Frequency method per group 1 Vehicle control
group 0 Single Intragastric 5 administration administration 2
Positive compound 10 mg/kg Single Intragastric 5 (Sotagliflozin)
administration administration 4 WXD010 10 mg/kg Single Intragastric
5 administration administration
[0217] Experiment Procedure:
[0218] 1. Animal Adaptation and Preparation:
[0219] The experimental animals were allowed to adapt to the
environment in the animal room for one week after they arrived.
[0220] 2. Fasting and Drug Administration.
[0221] The animals fasted in the metabolic cage for 18 hours, then
were given drugs or vehicle (2 ml/kg) according to the above table,
and subsequently were immediately given 50% glucose solution (2
g/kg, 4 ml/kg).
[0222] 3. Urine glucose and blood glucose test.
[0223] 2 hours after the administration of glucose solution, feed
intake was restored. Blood samples were taken at 0 min, 15 min, 30
min, 45 min, 60 min and 120 min respectively, and urine samples
were collected at 0-24 h to test blood glucose, urine glucose
(mg/200 g) and urine volume respectively.
[0224] 4. Data Analysis:
[0225] All values are represented as averages. Statistical analysis
was performed using Graphpad Prism 6 one-way analysis of variance
and Tukey's multiple comparison test. P value of less than 0.05 is
considered statistically significant.
[0226] The experimental results are shown in Table 8:
TABLE-US-00015 TABLE 8 Results of glucose tolerance test in rats.
Vehicle Positive control compound Compound group (Sotagliflozin)
WXD010 OGTT blood 1134 790**** 720**** glucose level AUC.sub.0-2 hr
(mol/L .times. min) Urine glucose 0.8 2843.2**** 2118.7**** level
(mg/200 g, BW) Urine volume 11.5 26.4**** 20.0**** (mL/200 g, BW)
Notes: *p < 0.05, **p < 0.01, **p < 0.001, ****p <
0.0001 vs. vehicle control group.
Conclusion: Compared with the Vehicle Control Group, the Compound
of the Present Disclosure can Significantly Reduce the Blood
Glucose AUC Level within 2 Hours and Increase the 24-Hour Urine
Glucose Excretion Level of Animals. Compared with the Positive
Compound, the Compound of the Present Disclosure has a Lower Level
of Urine Glucose Under the Same Hypoglycemic Effect, which is
Helpful to Reduce the Side Effects of Urinary Tract Infection
Experimental Example 4: Study on the In Vivo Pharmacodynamics in
Diabetic Db/Db Mice
[0227] Summary of the Experiment
[0228] 1. Animal Information:
TABLE-US-00016 Species db/db mice Grade SPF animal Age 5 weeks old
The age at which the experiment 6 weeks old started Body weight
range about 25 g Gender Male Supplier Model Animal Research Center
of Nanjing University Supplier address Nanjing, Jiangsu, China
[0229] 2 Animal Feeding
[0230] The animals were kept in an animal breeding room with
strictly controlled environmental conditions after they arrived.
The animal breeding room was maintained at a temperature of
20-24.degree. C. and a humidity of 40-70%. The temperature and
humidity in the feeding room were monitored in real time by a
hygrothermograph, and the temperature and humidity were recorded
twice daily (one in the morning and the other in the afternoon).
The lighting of the animal feeding room was controlled by an on-off
electronic timing system, with lights for 12 hours and dark for 12
hours daily (turned on at 7:00 a.m. and turned off at 19:00 p.m).
The mice were raised in separate cages and were given free access
to feed (reproductive feed 17053113 for rats and mice, available
from Beijing Keao Xieli Feed Co., Ltd.) and water during the
experiment.
[0231] 3. Experiment Grouping:
TABLE-US-00017 Administration Number of Administration method and
animals Group Treatment cycle @ dose frequency per group 1 Vehicle
Weeks 1-4 @0 mg/kg Intragastric 6 control Weeks 5-8 @0 mg/kg
administration, group once daily 2 WXD003 Weeks 1-4 @5 mg/kg
Intragastric 6 Weeks 5-8 @10 mg/kg administration, once daily 3
WXD010 Weeks 1-4 @5 mg/kg Intragastric 6 Weeks 5-8@10 mg/kg
administration, once daily
Experiment Procedure
[0232] 1. Administration of Drug
[0233] During the experiment, the animals were administrated with
the corresponding vehicles or drugs according to the group with an
administration time of 16:00 and an administration period of 8
weeks.
[0234] The dose was 5 mg/kg from week 1 to week 4; and the dose was
10 mg/kg from week 5 to week 8.
[0235] 2. Blood Glucose Level
[0236] Random and fasting blood glucose levels were measured once a
week.
[0237] The random blood glucose level was measured at 10:00
a.m.
[0238] Fasting blood glucose test: The mice fasted from 10:00 a.m.,
and blood glucose level was firstly measured at 16:00. Then the
mice were administered with the drugs, and 2 hours later blood
glucose level was measured again, and then feed intake was
restored.
[0239] 3. Oral Glucose Tolerance Test (OGTT).
[0240] At the end of the experiment (i.e. the last 3 days of
administration), the animals fasted for 6 hours, then were given a
single administration of glucose aqueous solution at a dose of 2
g/kg. The time of the glucose administration was recorded as 0 min.
Blood glucose levels of the animals were detected at 0 min before
the glucose administration, and 15 min, 30 min, 90 min and 120 min
after the glucose administration, respectively. The glucose
tolerance curve was drawn according to the data of blood glucose
levels vs time, and the area under the curve (AUC) was calculated.
The administration was given at 16:00.
[0241] 4. Biochemical Detection
[0242] At weeks 4 and 8 of the experiment, the animals fasted for 6
hours, and blood samples were collected to measure glycosylated
hemoglobin.
[0243] 5. Body Weight and Food Consumption
[0244] During the experiment, the body weight of the animals was
monitored once daily, and food consumption was monitored twice a
week.
[0245] 6. Data Processing and Analysis
[0246] All of the data were entered into an Excel document and
expressed in the form of mean.+-.S.E.M. The differences between
groups were compared using graphpad Prism 6 software and one-way
analysis of variance (ANOVA). P value of less than 0.05 is
considered a significant difference.
[0247] The results of random blood glucose experiment from week 1
to week 8 are shown in Table 9:
TABLE-US-00018 TABLE 9 Results of random blood glucose experiment
Vehicle control Group group WXD003 WXD010 Week 1 27.4 .+-. 2.26
14.3 .+-. 2.28 *** 16.7 .+-. 1.00 ** Week 2 21.8 .+-. 2.09 15.7
.+-. 0.88 13.5 .+-. 2.46 * Week 3 25.6 .+-. 2.65 13.6 .+-. 1.40 ***
14.1 .+-. 0.97 *** Week 4 27.4 .+-. 3.33 13.3 .+-. 0.81 **** 15.4
.+-. 1.34 *** Week 5 26.9 .+-. 3.67 14.1 .+-. 1.28 ** 13.2 .+-.
1.09 ** Week 6 28.8 .+-. 1.84 10.1 .+-. 1.26 **** 10.6 .+-. 1.48
**** Week 7 27.2 .+-. 2.63 12.5 .+-. 1.23 **** 9.8 .+-. 0.82 ****
Week 8 27.6 .+-. 2.96 14.4 .+-. 1.77 *** 9.2 .+-. 0.68 **** Notes:
* p < 0.05, ** p < 0.01, ** p < 0.001, **** p < 0.0001
vs. vehicle control group.
Conclusion: Compared with the Vehicle Control Group, the Compound
of the Present Disclosure can Reduce the Random Blood Glucose Level
of Animals; the Compound of the Present Disclosure can Further
Reduce the Random Blood Glucose Level of Animals with the Increase
of Dose
[0248] The results of fasting blood glucose test from week 1 to
week 8 are shown in Table 10:
TABLE-US-00019 TABLE 10 Results of fasting blood glucose test
Vehicle control Group group WXD003 WXD010 Week 1 21.8 .+-. 3.30
14.8 .+-. 2.34 14.7 .+-. 2.50 Week 2 26.8 .+-. 3.26 19.9 .+-. 1.32
14.4 .+-. 1.47 *** Week 3 28.7 .+-. 3.00 15.8 .+-. 1.48 ** 15.8
.+-. 1.40 ** Week 4 28.9 .+-. 3.35 14.3 .+-. 1.29 *** 14.9 .+-.
1.24 *** Week 5 25.7 .+-. 2.82 17.4 .+-. 1.72 * 15.9 .+-. 0.79 *
Week 6 28.5 .+-. 3.15 10.9 .+-. 1.42 **** 11.0 .+-. 1.67 **** Week
7 30.0 .+-. 2.58 11.7 .+-. 1.30 **** 9.6 .+-. 0.91 **** Week 8 31.4
.+-. 1.86 13.0 .+-. 1.36 **** 8.1 .+-. 0.51 **** Notes: * p <
0.05, ** p < 0.01, ** p < 0.001, **** p < 0.0001 vs.
vehicle control group.
Conclusion: Compared with the Vehicle Control Group, the Compound
of the Present Disclosure can Significantly Reduce the Fasting
Blood Glucose Level of Animals; by Increasing the Dose, the
Compound of the Present Disclosure can Further Reduce the Fasting
Blood Glucose Level of Animals
[0249] The results of oral glucose tolerance test (OGTT) at week 8
are shown in Table 11:
TABLE-US-00020 TABLE 11 Results of oral glucose tolerance test
(OGTT) at week 8 Vehicle control Compound group WXD003 WXD010 OGTT
blood 4515.5 .+-. 160.22 2806.5 .+-. 155.12**** 2118.4 .+-.
99.17**** glucose level AUC.sub.0-2 hr (mol/L .times. min) Notes:
*p < 0.05, **p < 0.01, **p < 0.001, ****p < 0.0001 vs.
vehicle control group.
Conclusion: Compared with the Vehicle Control Group, the Compound
of the Present Disclosure can Significantly Reduce the Blood
Glucose Level AUC within 2 Hours (AUC.sub.0-2hr) of Animals
[0250] The results of glycosylated hemoglobin (HbA1c) test at weeks
4 and 8 are shown in Table 12:
TABLE-US-00021 TABLE 12 Results of glycosylated hemoglobin (HbA1c)
at weeks 4 and 8 Week 4 Week 8 Rate of Rate of Decline of Decline
of Group HbA1c (%) HbA1c (%) HbA1c (%) HbA1c (%) Vehicle 8.4 0 9.6
0 control group WXD003 5.7*** -32 5.9**** -39 WXD010 5.8*** -31
5.1**** -47 Notes: *p < 0.05, **p < 0.01, **p < 0.001,
****p < 0.0001 vs. vehicle control group.
Conclusion: Compared with the Vehicle Control Group, the Compound
of the Present Disclosure can Significantly Reduce the Level of
Glycosylated Hemoglobin (HbA1c) of Animals; the Compound of the
Present Disclosure can Further Reduce the Level of Glycosylated
Hemoglobin (HbA1c) of Animals with the Increase of Dose
[0251] The results of body weight and food consumption are shown in
FIGS. 1 and 2. It is concluded that after 8 weeks of
administration, compared with the vehicle control group, the
animals in the administration group did not show significant change
in the body weight and food consumption, indicating that the
animals have good tolerance to the compound of the present
disclosure.
* * * * *