CN108383801A - The inhibitor of SGLT2 albumen and application - Google Patents

The inhibitor of SGLT2 albumen and application Download PDF

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CN108383801A
CN108383801A CN201810237106.1A CN201810237106A CN108383801A CN 108383801 A CN108383801 A CN 108383801A CN 201810237106 A CN201810237106 A CN 201810237106A CN 108383801 A CN108383801 A CN 108383801A
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compound
sglt2
inhibitor
stood alone
albumen
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CN108383801B (en
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于磊
徐巧枝
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Abstract

The screening technique of the inhibitor of SGLT2 albumen a kind of and compound in the application and inhibitor for preparing the drug for the treatment of diabetes.Wherein, the inhibitor includes the compound with structure shown in formula (1)：The inhibitor can be used as active primer treatment diabetes.

Description

The inhibitor of SGLT2 albumen and application
Technical field
The present invention relates to the relevant field for the treatment of diabetes, further to a kind of SGLT2 (Na+-glucose Cotransporter 2, OMIM：* 182381, also referred to as SLC5A2) albumen inhibitor, further relate to a kind of compound and treating The application of diabetes further relates to the screening technique of SGLT2 protein inhibitors.
Background technology
Kidney plays an important roll in adjusting blood glucose level, and there are two class glucose transporter in kidney, wherein one Class is the glucose transporter (Na+-glucose cotransporter, SGLT) that sodium relies on, and consumption energy is with active mode Inverse concentration gradient transports glucose.Wells etc. is found that a kind of referred to as SGLT2 (OMIM from the cDNA library of people's kidney:* 182381, be also SLC5A2) transport protein.
Familial renal glucosuria (Familial renal Glucosuria, FRG) is with fasting blood-glucose and carbohydrate tolerance test Normally, the disease lowered with the proximal tubular glucose re-absorption function that lasting glucose in urine is main feature, and patient is mostly Without renal tubular function and the exception of structure, prognosis bona.In the prior art in familial renal glucosuria patient's nephridial tissue Expression study has been carried out, it is found that wild type and saltant type SGLT2 albumen are expressed in renal cells, wherein wild type SGLT2 can be expressed normally, and abnormal distribution and expression occur on cell membrane in SGLT2 muteins, to again It is determined that familial renal glucosuria should be defined as codominant inheritance, and the incomplete penetrance previously thought considers and the open countries SGLT2 Raw type albumen it is compensatory related.
Research in recent years finds that the expression of the SGLT2 of diabetic and function are obviously raised, and familial renal glucosuria is suffered from Person prognosis bona, it is therefore, special to block proximal convoluted tubule SGLT2 transport proteins, and then inhibit glucose reabsorption to control blood glucose Level becomes the hot spot studied at present.Phloridzin (phlorizin) is that french chemist in 1835 is detached from apple bark The first SGLT inhibitor gone out finds that phloridzin can reduce blood glucose in subsequent many diabetic animals research, restores The sensibility of insulin, but since oral administration biaavailability is relatively low, specificity is not high, and metabolite may inhibit The function of the albumen such as GLUT1 has some side effects, so being not applied to glycosuria in central nervous system and other systems The clinical treatment of disease.
Invention content
(1) technical problems to be solved
In view of this, the purpose of the present invention is to provide a kind of inhibitor of SGLT2 albumen and compound to be controlled in preparation The application of the drug of diabetes and the screening technique of SGLT2 protein inhibitors are treated, to solve at least one of above-described skill Art problem.
(2) technical solution
According to an aspect of the present invention, a kind of inhibitor of SGLT2 albumen is provided, includes with structure shown in formula (1) Compound：
Formula (1),
Wherein, A is stood alone as：
B is stood alone as：
According to another aspect of the invention, a kind of application of compound in the drug for preparing treatment diabetes is provided, In, the compound has structure shown in above formula (1)：
In accordance with a further aspect of the present invention, a kind of screening technique of SGLT2 protein inhibitors is provided, including：
SGLT2 albumen and compound is arranged docks condition；
Virtual screening is carried out in the database according to docking condition；
It is carried out, to further screening and the marking of virtual screening result, obtaining the change of setting score or more according to quasi-medicated property principle Close lead compound of the object as treating diabetes；
Wherein, setting SGLT2 albumen includes with the condition of docking of compound：
The effect for setting compound and SGLT2 is maintained by strong interaction by two parts, is on the one hand hydrophily The amino acid residue of stronger glycosyl part and pocket part outwardly forms more stable hydrogen bond；On the other hand it is small molecule aromatic rings Part and the hydrophobic interaction and π-π of the hydrophobic pocket of pocket higher depth interact；The lipid of compound Logarithm logP be -2~5, molecular weight be 200~500, hydrogen-bond donor 0~5, hydrogen bond receptor 0~10, rotatable number of keys are 0 ~10, heavy atom quantity is 20~70 and charge value is -2~2.
(3) advantageous effect
Through the above scheme, it is known that the beneficial effects of the present invention are：
Compound in the inhibitor of SGLT2 albumen through the invention can be used as active lead compound treatment sugar Urine disease.
Compound in the inhibitor of the SGLT2 albumen of the present invention includes hydrophilic radical and hydrophobic pocket, with SGLT2 albumen The polar pocket of active site, pocket waterborne interaction, inhibit the expression of SGLT2 albumen.
The compound of the present invention can be applied to prepare diabetes medicament.
Description of the drawings
Fig. 1 is the compound of the inhibitor of SGLT2 albumen of the embodiment of the present invention.
Fig. 2 is the compound and SGLT2 albumen action principle figures of inhibitor of the embodiment of the present invention.
Fig. 3 is the mutational site found in familial renal glucosuria patient of the embodiment of the present invention.
Fig. 4 is the structure-activity relationship figure of compound of the embodiment of the present invention and SGLT2 albumen.
Fig. 5 is the standard curve in specific example 1 of the present invention.
Specific implementation mode
To make the objectives, technical solutions, and advantages of the present invention clearer, below in conjunction with specific embodiment, and reference Attached drawing, the present invention is described in further detail.
Since the SGLT2 expression and functional study of the past in the prior art are only found that 2 kinds of mechanism therein, the application hair A person of good sense has found that remaining 4 kinds of mechanism plays an important roll in familial renal glucosuria pathogenic process, and specifies relevant bits Point.C.294C inventor is further discovered that>A/p.(F98L),c.736C>T/p. (P246S) is diabetes medicament (SGLT2 albumen Inhibitor) screening desired target point.Due to being still not very clear that medicament research and development is all sought to the exact 3D structures of SGLT2 The derivative or the like for looking for phloridzin, may be to other glucose transport proteins while inhibiting SGLT2 protein functions Different degrees of influence is also resulted in vain, produces ill-effect, and certain difficulty is caused in the R＆D process of drug.
One side according to the ... of the embodiment of the present invention provides a kind of inhibitor of SGLT2 albumen, including following compound：
Formula (1), wherein A is stood alone as：
B is stood alone as：
The compound of formula (1) as shown in Figure 1 is the docking structure of three small molecules, shown referring to Fig.1 comprising left and right The hydrophilic radical and hydrophobic grouping (mark in dashed rectangle) of both sides, and intermediate coupling part (dashed middle line box), (knot It closes shown in Fig. 2, for SGLT2 as target albumen, active site has apparent polar pocket (such as residue Ser460 in Fig. 2) With hydrophobic pocket (such as Trp289).
In Fig. 2, the combination of these compounds and SGLT2 mainly have three parts：1, the hydrophilic radical of compound with The residues such as Ser460 form interaction of hydrogen bond；2, the coupling part of hydrophilic radical and hydrophobic grouping is formed by residue Lys154 Hydrogen bond action carrys out the combination of stable compound；3, the hydrophobic grouping of compound enters hydrophobic pocket, forms hydrophobic effect；Especially when When hydrophobic grouping is aromatic rings, stronger pi-pi bond is formed with Trp289 residues, L286V mutation in addition can make the benzene first of small molecule Base portion point (structure in circle in Fig. 4) has enough spatial positions and protein-interacting.
It by above-mentioned inhibitor, can specifically inhibit SGLT2 protein functions, effectively reduce empty stomach and postprandial blood Sugar becomes the new way for the treatment of diabetes.
Above-mentioned inhibitor can do the isostructural drug of slabbing using powder as raw material.
In a further embodiment, the A can be stood alone as Preferably, A can be stood alone asPreferred A can be stood alone as
In some embodiments, the B is stood alone as Preferably, B is stood alone as Preferred B is stood alone as
The embodiment of the present invention also provides a kind of application of compound in the drug for preparing treatment diabetes, wherein described Compound has structure shown in formula (1) described above.
Another aspect according to the ... of the embodiment of the present invention provides a kind of screening technique of SGLT2 protein inhibitors, including：
SGLT2 albumen and compound is arranged docks condition；
Virtual screening is carried out in the database according to docking condition；
It is carried out, to further screening and the marking of virtual screening result, obtaining the compound conduct of high score according to quasi-medicated property principle The lead compound for the treatment of diabetes.
In the embodiment of the present invention, it is with the SGLT2 gene mutations found in the familial renal glucosuria patient of Inner Mongolia Basis, further to SGLT2 from gene mutation to protein expression, function progress in-depth study, for disclosing SGLT2 in family Property renal glucosuria pathogenesis in key effect and its mechanism of action clinical and basic research evidence and theoretical foundation (ginseng are provided As shown in Figure 3), by virtual screening, according to quasi-medicated property principle, we eliminate part the selection result, finally obtain small molecule Compound.By the interaction relationship of small molecule and receptor protein, we analyze and select the biology that small molecule can be used for the later stage Experimental verification.It is design, screening and the novel drugs research and development of Remedies for diabetes further by cell and animal level verification Significant target and important clue and foundation are provided.
Screening process includes：1, drug virtual screening：The target spot that drug screening is determined using the result of functional study, into one Walk the drug of screening treatment diabetes.SGLT2 albumen is carried out based on known antagonist phloridzin and its derivative or the like The Inhibition test of function as a result, establish quantitative structure activity relationship model, using the method for three-dimensional pharmacophore model search, changing It closes and carries out void in object database (commercial micromolecular compound library ACD-SC, MDDR and China Natural Product Database CNPD) Quasi- screening, finds active lead compound.And further screened according to quasi-medicated property principle, it gives a mark, finally obtains several Lead compound of a high score compound as treating diabetes, new clue and foundation are provided for medicament research and development.2, virtual sieve Select scheme：It is tested by the flexible docking of albumen and three drug molecules, we tentatively can be determined that：Patent medicine small molecule and albumen Effect maintained compared with strong interaction possibly via two parts, be on the one hand the stronger glycosyl part of hydrophily and pocket court The amino acid residue of outer portion forms more stable hydrogen bond；On the other hand it is small molecule aromatic moiety hydrophobic with pocket higher depth The hydrophobic interaction of pocket and π-π interactions.Virtual screening is done according to the docking condition set, in being calculated with Beijing The virtual screening platform of the heart is screened, and the small molecule libraries selected are patent medicine database and can provisioning database C.3, pharmacophore sieves Process is selected, is specifically included：Small molecule is selected：A/ small molecules have the general character of >=4 drug molecules, and the pharmacophore built out can be complete Number sieve goes out 5 small molecules；B/ small molecules have specificity difference recognition site to SGLT2 and SGLT1.Pharmacophore is established and test, 5 drugs can be filtered out；One decoy test group is set again, including 5 small molecules, it is optimal that complete 5 drugs are sifted out in test.
Pharmacophore screen marketed drug library all and can the docking top30% virtual screenings of giving a mark of provisioning database c work.
The higher small molecule of pharmacophore marking is selected, the specificity combined with amino acid residue according to visualization small molecule is chosen Select the small molecule of the embodiment of the present invention.
Fig. 4 is the structure-activity relationship figure of compound of the embodiment of the present invention and SGLT2 albumen.By existing three drug molecules pair It is connected in SGLT2 albumen, it is found that the docking result of three small molecules has certain rule.As shown in figure 4, from albumen Film inner region sees that the glycosyl part of three small molecules is overlapping, and small molecule is in the hydrophobic of pocket higher depth in addition to albumen depths Qu Jun has multiple hydrophobic functional groups.Scale model shows that SGLT2 and SGLT1 has the amino acid residue of mutation in pocket locations, Preliminary analysis finds that it may be because small molecule is more easy to be formed with hydrophobic pocket that three drug specificities, which are attached to SGLT2 pockets, Interaction of hydrogen bond, in addition L286V mutation can make the phenylmethyl moiety of small molecule have enough spatial positions and albumen phase interaction With (referring to Fig. 4 encircled portions).
Specific example 1 will be enumerated below, the effect of the compound of the embodiment of the present invention is verified in cellular level, find glycosuria The lead compound of disease treatment, method are as follows：
Cellular level is verified：
One, laboratory apparatus and reagent
Two, experimental method
1., cell prepare
1. with 0.25% trypsin digestion and cell, cell suspension is prepared, adjustment cell density is 2 × 105A/ml.
2. inoculating cell suspension (500 holes μ L/) in 24 orifice plates, by culture plate put culture in the incubator (37 DEG C, 5% CO2) overnight.
3. it is thin to form transfected plasmids by transfection reagent P-transter for selectivity plus transfected plasmids (SGLT-2 or GFP) Born of the same parents.
4. culture plate is put culture (37 DEG C of culture environment, 5%CO in the incubator2)48h。
2., drug-treated
1. after 48h, changing free serum culture into and being based on cultivating (culture environment in incubator：37 DEG C, 5%CO2), overnight.
2. being changed to serum-free sugar-free culture-medium, corresponding inhibitor can be selectively added, in the incubator culture (training Support environment：37 DEG C, 5%CO2) 40min.Here inhibitor has structure shown in formula (1)： Formula (1), A areB isThe inhibitor is labeled as T.
3. plus 2-DG (2-deoxyglucose)
1. 20 μ l 2-DG are added per hole.It is incubated that (condition is in incubator：37 DEG C, 5%CO2) 20min.
2. removing culture medium, washed 3 times with the PBS of precooling.
4. plus 2-DG
1. preparing lysis buffer：Add 1%Triton X-100 in NADP Extraction buffer (extraction bufferings Liquid) in.Prepare 220 μ l per hole.
2. standard curve：
A. prepare 5 μM of 2-DG6P (2-deoxyglucose 6-phosphate)：5 μ l 5mM Standard (marks are added Quasi- product) in 4995 μ l distilled water, mixing.
B. each master sample dilution process enters the following table 1
Table 1
Serial number Premix+H2O 2-DG6P(μM)
1 100μl+0μl 5.0
2 60μl+40μl 3.0
3 30μl+70μl 1.5
4 0μl+100μl 0
3. 200 μ l lysis buffers are added in the hole of 24 orifice plates, shaking table shakes 5min.
4. 50 μ l cell pyrolysis liquids samples and 50 μ l master samples are drawn respectively to manage in 1.5ml EP, in 80 DEG C, 10min.
5. 50 μ l NADPH extraction buffer (Extraction buffer) are added in cell sample pipe, 50 μ l are added H2O is in master sample pipe, -20 DEG C, 5min.
6. preparing WR1：Add 10 μ l Assay buffer (reaction buffer), 1 μ l G6PDH (glucose 6- Phosphate dehydrogenase) Enzyme, in 1 μ l NADP.Often pipe prepares 12 μ l.Often 10 μ l WR1 are added in pipe, in 37 DEG C, 60min.
7. often 50 μ l NADPH extraction buffer, in 80 DEG C, 15min are added in pipe.
8. often pipe is added 50 μ l NADP extraction buffer, -20 DEG C, 5min.
9. 50 μ l are respectively drawn in cell sample and master sample, in the hole of 96 orifice plate of black.
10. preparing WR2：Add 45 μ l Assay buffer, 1 μ l Enzyme A, 1 μ l Enzyme B, 10 μ l G6P (glucose 6-phosphate) Reagent, 5 μ l Probe.
11. 50 μ l WR2 are added per hole, 0min and 20min light absorption values are measured respectively at 530/585.
Three, experiment packet
Experiment packet situation such as the following table 2, each group is done three multiple holes.
Table 2
Group Details Drug concentration
C1 Transfected plasmids (GFP) cell
C2 Transfected plasmids (SGLT-2) cell
C3 Normal cell
C4 Transfected plasmids (SGLT-2) cell+solvent
T Transfected plasmids (SGLT-2) cell+inhibitor T 1mM
Four, experimental result and analysis
1, standard curve
Table 3
0(μM) 1.5(μM) 3(μM) 5(μM)
0min 13286 18528 20563 24000
20min 14764 24054 31384 42060
20-0min 1478 5526 10821 18060
Upper table is light absorption value as a result, linear standard curve shown in fig. 5 can be fitted to.
2. glucose uptake data analysis
4 Tukey multiple comparative tests of table
Tukey Multiple range tests verification shown in table 4 is carried out to the glucose uptake amount of group C1-C4 and T, can be seen It is other higher than C1-C4 groups to go out the other glucose uptake amount of T groups, it, can be as sugar with preferably with preferable inhibition The lead drug of urine disease treatment.
Particular embodiments described above has carried out further in detail the purpose of the present invention, technical solution and advantageous effect Describe in detail bright, it should be understood that the above is only a specific embodiment of the present invention, is not intended to restrict the invention, it is all Within the spirit and principles in the present invention, any modification, equivalent substitution, improvement and etc. done should be included in the protection of the present invention Within the scope of.

Claims (10)

1. a kind of inhibitor of SGLT2 albumen, it is characterised in that include the compound with structure shown in formula (1)：
Wherein, A is stood alone as：
B is stood alone as：
2. inhibitor according to claim 1, which is characterized in that the A is stood alone as
3. inhibitor according to claim 1, which is characterized in that the B is stood alone as
4. inhibitor according to claim 2, which is characterized in that the A is stood alone as
5. inhibitor according to claim 3, which is characterized in that the B is stood alone as：
6. inhibitor according to claim 1, which is characterized in that the A isThe B is
7. a kind of application of compound in the drug for preparing treatment diabetes, wherein the compound has knot shown in formula (1) Structure：
Wherein, A is stood alone as：
B is stood alone as：
8. application according to claim 7, which is characterized in that the compound is used to inhibit the expression of SGLT2 albumen, institute It is I types and type-2 diabetes mellitus to state diabetes.
9. a kind of screening technique of SGLT2 protein inhibitors, which is characterized in that including：
SGLT2 albumen and compound is arranged docks condition；
Virtual screening is carried out in the database according to docking condition；
It is carried out, to further screening and the marking of virtual screening result, obtaining the compound of setting score or more according to quasi-medicated property principle Lead compound as treating diabetes；
Wherein, setting SGLT2 albumen includes with the condition of docking of compound：
The effect for setting compound and SGLT2 is maintained by strong interaction by two parts, is on the one hand that hydrophily is stronger The amino acid residue of glycosyl part and pocket part outwardly form more stable hydrogen bond；On the other hand it is small molecule aromatic moiety It interacts with the hydrophobic interaction and π-π of the hydrophobic pocket of pocket higher depth；The logarithm of the lipid of compound Value logP be -2~5, molecular weight be 200~500, hydrogen-bond donor 0~5, hydrogen bond receptor 0~10, rotatable number of keys be 0~ 10, heavy atom quantity is 20~70 and charge value is -2~2.
10. screening technique according to claim 9, which is characterized in that carried out to virtual screening knot according to quasi-medicated property principle Fruit further screening and marking in the condition further screened include：
There is the small molecule selected the general character of >=4 drug molecules, the pharmacophore built out can sift out 5 small molecules totally；
Small molecule has specificity difference recognition site to SGLT2 and SGLT1.
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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004103980A1 (en) * 2003-05-21 2004-12-02 Biovitrum Ab Inhibitors of 11-beta-hydroxy steroid dehydrogenase type i
CN1688597A (en) * 2002-08-23 2005-10-26 橘生药品工业株式会社 Pyrazole derivatives, medicinal composition containing the same, medicinal use thereof, and intermediate for production thereof
CN105193811A (en) * 2014-06-25 2015-12-30 中国药科大学 SGLT2 inhibitor and application thereof
EP2998295A1 (en) * 2014-09-16 2016-03-23 Sanofi Aryl substituted naphthyl sulfonamide derivatives as keap-1 modulators for the treatment of diabetes, obesity, dyslipidemia and related disorders
CN107001283A (en) * 2014-07-31 2017-08-01 国家健康与医学研究院 Flt3 receptor antagonists
CN107155325A (en) * 2014-08-27 2017-09-12 Cj医药健康株式会社 Novel amino phenyl sulfonyl acetic ester derivative and application thereof

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1688597A (en) * 2002-08-23 2005-10-26 橘生药品工业株式会社 Pyrazole derivatives, medicinal composition containing the same, medicinal use thereof, and intermediate for production thereof
WO2004103980A1 (en) * 2003-05-21 2004-12-02 Biovitrum Ab Inhibitors of 11-beta-hydroxy steroid dehydrogenase type i
CN105193811A (en) * 2014-06-25 2015-12-30 中国药科大学 SGLT2 inhibitor and application thereof
CN107001283A (en) * 2014-07-31 2017-08-01 国家健康与医学研究院 Flt3 receptor antagonists
CN107155325A (en) * 2014-08-27 2017-09-12 Cj医药健康株式会社 Novel amino phenyl sulfonyl acetic ester derivative and application thereof
EP2998295A1 (en) * 2014-09-16 2016-03-23 Sanofi Aryl substituted naphthyl sulfonamide derivatives as keap-1 modulators for the treatment of diabetes, obesity, dyslipidemia and related disorders

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
仇缀百: "《药物设计学》", 30 June 2008, 浙江大学出版社 *
尤启冬: "《药物化学》", 31 August 2011, 人民卫生出版社 *

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