CN110423267A - A kind of cyclic ester peptides Gq protein inhibitor and its preparation method and application - Google Patents
A kind of cyclic ester peptides Gq protein inhibitor and its preparation method and application Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
- A61P11/06—Antiasthmatics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Abstract
The invention belongs to pharmaceutical technology field, in particular to a kind of cyclic ester peptides Gq protein inhibitor and its preparation method and application.Cyclic ester peptides Gq protein inhibitor, specially as follows shown in logical formula (I).Test result is shown, selective Gq protein inhibitor as a kind of structure novel, compared with YM-254890 and FR900359, compound in the present invention has the advantages that synthesis is relatively simple and metastable, most of such cyclic ester peptides have certain Gq inhibitory activity and selectivity, have exploitation at the potentiality of new and effective Gq protein selectivity inhibitor.
Description
Technical field
The invention belongs to pharmaceutical technology field, in particular to a kind of cyclic ester peptides Gq protein inhibitor and preparation method thereof and
Using.
Background technique
GPCR and G-protein in vivo can be by mediating a plurality of signal transduction pathway to control a series of intracorporal lifes of people
Function is managed, such as in cancer, GPCR and G-protein can pass through the development of complicated signal path net influence tumour.GPCR connects
After receiving the extracellular information such as neurotransmitter, hormone, G protein activation and then interacting with G-protein, so that outer signals be passed
It is handed to intracellular.Since G-protein takes part in a series of signal transductive process, thus finds one and suitably directly act on G egg
White agonist or inhibitor has very big researching value.
YM-254890 is that only two specificities being currently known inhibit one of the G-protein inhibitor of Gq albumen, thus
YM-254890 is the priceless tool molecule of a research G-protein activation and the protein mediated cell effect of Gq.YM-254890
By combining the hydrophobic cavity of Gq albumen, stablizes the non-live condition conformation of Gq protein binding GDP, inhibit Gq albumen living to play
The effect of change.Research shows that YM-254890 has extensive physiological activity, the platelet aggregation induced by ADP can inhibit, this
Also there is antithrombotic and thrombolysis activity in external mouse model.2017, Michaela Matthey etc. will also be by YM-
254890 analogue FR900359 is applied to obstructive lung disease research aspect.It was found that compound FR900359 can effectively press down
The Gq protein signal of the airway smooth muscle cells of mouse processed and people, in addition, FR900359 may also suppress bronchoconstriction reaction, and
Cause lasting respiratory tract relaxation in the airway tissue of mouse, pig and people.Research is it has also been found that the wild-type mice of health is inhaled
It will lead to compound intrapulmonary local concentration raising after entering FR900359, can prevent respiratory tract from shrinking without to blood pressure and heart rate
Deng generation acute hazard, furthermore it has also been found that FR900359 can prevent Respiratory insufficiency on mouse model.Result of study table
It is bright FR900359 be can be used as into a kind of selective Gq protein inhibitor to be applied to internal respiratory tract, in the pulmonary diseases such as asthma
It may be to realize a kind of bronchiectasic effective therapeutic strategy.However, as a kind of effective Gq protein inhibitor, YM-
254890 synthesis is more difficult, and the presence of dehydroalanine structure largely reduces the stabilization of the cyclic ester peptide.
Property.
Based on this, it is necessary to develop new selective Gq protein inhibitor for this field.
Summary of the invention
In order to overcome the shortcomings and deficiencies of the prior art described above, the primary purpose of the present invention is that providing a kind of cyclic ester peptides
Gq protein inhibitor.
Another object of the present invention is to provide the preparation method of above-mentioned cyclic ester peptides Gq protein inhibitor.
Still a further object of the present invention be to provide above-mentioned cyclic ester peptides Gq protein inhibitor prepare it is antitumor, antithrombotic and
It is applied in anti-asthmatic medicament.
The purpose of the present invention is realized by following proposal:
A kind of cyclic ester peptides Gq protein inhibitor specially such as leads to formula (I) compound represented or its is pharmaceutically acceptable
Salt or ester or solvate:
Wherein,
N is selected from 0,1 or 2；
Y is selected from O, S, Se ,-CH2,-NH- ,-CONH- ,-NHCO- ,-COO- or-OOC-；
Z is selected from benzyl, aryl, heteroaryl, naphthenic base, Heterocyclylalkyl, straight chained alkyl, guanidine radicals, phosphate, naphthalene, quinoline
Base, benzofuranyl or indyl；Preferably, the heteroaryl and Heterocyclylalkyl independently can be selected from N, O or S containing 1~3
Hetero atom；It is highly preferred that Z can be replaced by 1~3 identical or different R；R be hydrogen, fluorine, chlorine, smell, iodine, hydroxyl, nitro,
At least one of cyano, trifluoromethyl, methoxyl group and trifluoromethoxy.
Preferably, the logical formula (I) compound represented has structure shown in I-1~I-33:
A kind of preparation method of above-mentioned cyclic ester peptides Gq protein inhibitor, specifically includes the following steps:
(1) by 2- chlorine trityl chloride resin (2-CTC resin), N- fluorenes methoxy carbonyl acyl group-N- methyl-L-alanine
(Fmoc-N-Me-Ala-OH) after mixing in DCM with n,N-diisopropylethylamine (DIEA), it is molten to add piperidines/DMF mixing
The reaction was continued obtains compound (A) for liquid；
(2) by N- fluorenes methoxy carbonyl acyl group-l-Alanine (Fmoc-Ala-OH), 2- (7- aoxidize benzotriazole)-N, N,
N', N'- tetramethylurea hexafluorophosphate (HATU), trimethylpyridine (collidine) are dissolved in after DMF and step (1) gainedization
Object (A) hybrid reaction is closed, piperidines/DMF mixed solution is added the reaction was continued and obtain compound (B)；
(3) by compound G, HATU and collidine be dissolved in after DMF with compound (B) hybrid reaction obtained by step (2),
Piperidines is added, the reaction was continued obtains compound (C)；Wherein the compound G general structure is as follows:
Wherein Z, Y, n all have meaning described in logical formula (I)；
(4) it is mixed after compound 16, HATU being dissolved in DMF with collidine with compound (C) obtained by addition step (3)
Reaction, then adds phenylsilane (PhSiH3)、Pd(PPh3)4The reaction was continued with DCM, obtains compound (D)；
(5) compound 25, HATU and collidine are dissolved in DMF, are then mixed instead with compound (D) obtained by step (4)
It answers, obtains compound (E)；
(6) by compound (E) and trifluoroacetic acid/tri isopropyl silane/methylene chloride (TFA/TIPS/ obtained by step (5)
DCM) after hybrid reaction, filtrate is filtered and collected, filtrate is concentrated to get compound (F), is i.e. I -1 chain precursor of cyclic ester peptide.；
(7) after compound (F) obtained by step (6) being dissolved in DMF, with DIEA and hexafluorophosphoric acid benzotriazole -1- base-oxygroup
Tripyrrole alkyl phosphorus (PyBop) hybrid reaction obtains the cyclic ester peptides Gq protein inhibitor as shown in formula (I).
Above-mentioned cyclic ester peptides Gq protein inhibitor is preparing antitumor, applies in antithrombotic and anti-asthmatic medicament.
The present invention also provides a kind of Pharmaceutical compositions comprising above-mentioned cyclic ester peptides Gq protein inhibitor and at least one
Pharmaceutic adjuvant.
The present invention compared with the existing technology, have the following advantages and the utility model has the advantages that
Gq protein inhibiting activity test result of the present invention shows that most of such cyclic ester peptides have certain Gq
Inhibitory activity and selectivity.As the selective Gq protein inhibitor of a kind of structure novel, with YM-254890 and FR900359 phase
Than the compound in the present invention has the advantages that synthesis is relatively simple and metastable, has exploitation at new and effective Gq egg
The potentiality of white selective depressant.
Specific embodiment
Below with reference to embodiment, the present invention is described in further detail, and embodiments of the present invention are not limited thereto.
Agents useful for same can routinely be bought unless otherwise specified from market in embodiment.
Room temperature described in embodiment is 15~35 DEG C.
The synthesis of embodiment 1:I-1
The preparation of compound 6
By compound 1 (N- fluorenylmethyloxycarbonyl-N- methyl O- tert-butyl-Serine) (1.00g, 2.52mmol) molten dichloro
It in methane (DCM, 10mL), is added trifluoroacetic acid (TFA, 3mL), 3h is to complete for room temperature reaction, is concentrated under reduced pressure to give compound 2.
By compound 2 (850mg, 2.48mmol) and NaHCO3(254mg, 3.02mmol) is dissolved in H2O (5mL) obtains solution A, separately by methyl
Trioctylmethylammonium chloride (aliquat336,1.12g, 2.77mmol) and bromopropene (Allyl-Br, 0.46g, 3.77mmol) are dissolved in
DCM (5mL) solution B, by solution B be added solution A in, room temperature reaction overnight, then add Allyl-Br (0.15g,
1.26mmol), room temperature the reaction was continued for 24 hours, then reaction solution is extracted with DCM (in triplicate 3, use 10mL every time),
Merge organic phase, then after being washed with saturated brine, passes through anhydrous Na2SO4It dries, filters, is concentrated under reduced pressure, silica gel column chromatography purifying
It obtains compound 3 (740mg, yield 77.2%).Again by compound 3 (740mg, 1.94mmol) and paratoluensulfonyl chloride (Ts-Cl,
1.85g, 9.71mmol) it is dissolved in pyridine (3mL), then after reaction overnight at 0 DEG C, diluted with ethyl acetate (EA, 20mL), then according to
It is secondary to use H2O (10mL), saturated lemon aqueous solution (being repeated 2 times, each 10mL), saturated sodium bicarbonate aqueous solution (10mL), satisfy
It is washed with salt water (10mL), organic phase passes through anhydrous Na2SO4It after drying, filters, is concentrated under reduced pressure, be purified by silica gel column chromatography to change
It closes object 4 (428mg, yield 41.2%).Selenophenol (PhSeOH 187mg, 1.19mmol) is dissolved in DMF (3mL), is cooled to 0 DEG C
Reaction solution is obtained, then NaOH (31.7mg, 0.79mmol) is dissolved in H2In O (1mL) and instill reaction solution, in 0 DEG C of reaction 5min,
Compound 4 (425mg, 0.79mmol) is dissolved in acetone (5mL) again and instills reaction solution, it is dilute with EA (50mL) after 0 DEG C of reaction 3h
It releases, successively with saturation NH4Cl aqueous solution (being repeated 3 times, each 20mL), saturated brine (20mL) washing, organic phase uses anhydrous
Na2SO4It is concentrated under reduced pressure after drying, filtering, silica gel column chromatography purifies to obtain compound 5 (165mg, yield 40.0%).Again by compound
5 (160mg, 0.31mmol) and tetrakis triphenylphosphine palladium [Pd (PPh3)4] it is placed in reaction flask, it is protected from light and is led to nitrogen protection, nothing is added
Morpholine (morpholine, 28.1 μ L, 0.32mmol) is separately dissolved in anhydrous THF (1.5mL) by water tetrahydrofuran (THF, 1.5mL)
After be slowly dropped into reaction flask, 30min is to complete for room temperature reaction, and EA (20mL) dilution, 2M HCL aqueous solution (is repeated 3 times, every time
It 10mL) washs, organic phase passes through anhydrous Na2SO4It is concentrated under reduced pressure after drying, filtering, silica gel column chromatography purifies to obtain compound 6
(110mg, 74.4%).
MS(ESI)calcd.for C25H23NO4Se+[M+H]+481.08,found 481.08。1H NMR(400MHz,
CDCl3) (2:1 rotamer ratio,#The more rotational isomer signal of content is represented, * represents the less rotational isomeric of content
Body signal) δ 7.78-7.69 (m, 2H), 7.61-7.43 (m, 4H), 7.42-7.27 (m, 4H), 7.24-7.18 (m, 3H), 4.65#
(dd, J=10.4,4.4Hz, 1H), 4.54* (dd, J=10.0,4.8Hz, 1H), 4.44-4 4.32 (m, 2H), 4.25#(t,J
=7.2Hz, 1H), 4.10* (t, J=6.0Hz, 1H), 3.52-3.43 (m, 1H), 3.36-3.22 (m, 1H), 2.89#(s,3H),
2.83*(s,3H).13C NMR(125MHz,CDCl3)δ173.58,155.66,142.81,140.29,132.38,132.18,
128.21,126.71,126.54,126.07,124.10,118.97,66.98#,66.79*,46.13#,46.05*,31.94,
28.68,25.66。
The preparation of compound 16
Weigh Compound 7 (3.01g, 18.05mmol) is dissolved in anhydrous DMF (15mL), and Cs is added2CO3(6.47g,
Suspension 19.86mmol) is obtained, 10min is then stirred at room temperature, then by suspension ice bath to 0 DEG C, addition cylite (BnBr,
2.2mL, 18.42mmol), in 0 DEG C of stirring 30min, reaction 2h is then stirred at room temperature to complete.By gained reaction solution EA
(100mL) dilution, is then filtered to remove white solid, acquired solution is successively used to saturated sodium bicarbonate aqueous solution (100mL), water
The washing of (being repeated 2 times, each 100mL), saturated brine (100mL), gained organic phase anhydrous Na2SO4It is depressurized after drying, filtering
Concentration, then purifies to obtain compound 8 (3.82g, 83.1%) by silica gel column chromatography.By compound 8 (3.31g, 12.95mmol)
DCM is dissolved in N- fluorenylmethoxycarb-nyl -nyl O-tert-butyl-L-threonine (Fmoc-Thr (tBu)-OH, 6.18g, 15.54mmol)
In (50mL), be added dicyclohexylcarbodiimide (DCC, 3.21g, 15.54mmol) and 4-dimethylaminopyridine (DMAP,
0.32g, 2.59mmol), 6h is stirred at room temperature to complete, is filtered to remove white solid, then acquired solution is concentrated under reduced pressure, then lead to
It crosses silica gel column chromatography and purifies to obtain compound 9 (6.65g, 81.1%).Compound 9 (3.22g, 5.04mmol) is dissolved in DCM
(20mL) is added diethylamine (8.0mL), and reaction 4h is stirred at room temperature to complete, gained reaction solution is concentrated under reduced pressure, silicon is then passed through
Plastic column chromatography obtains compound 10 (1.80g, 86.5%).Compound 10 (1.80g, 4.36mmol) is dissolved in DCM (15mL), ice
Bath is added triethylamine (1.8mL, 12.96mmol), then chloroacetic chloride (600 μ L, 8.44mmol) is added dropwise, and ice bath reacts 20min, moves
To room temperature reaction 2h to complete, obtained reaction solution is washed with saturated brine, then uses anhydrous Na2SO4After drying, filtering, by institute
After solution is concentrated under reduced pressure, compound 11 (1.30g, 65.7%) is purified to obtain by silica gel column chromatography.By compound 11
(1.30g, 2.86mmol) is dissolved in DCM (10mL), is added TFA (4mL), and reaction 2h is stirred at room temperature to complete, by gained reaction solution
It is concentrated under reduced pressure, compound 12 (0.95g, 83.3%) is then purified to obtain by silica gel column chromatography.By compound 12 (0.95g,
It 2.38mmol) is dissolved in DCM (10mL), N- [tertbutyloxycarbonyl]-N- methyl-O- methyl-L-threonine (Boc-N-Me-Thr is added
(Me)-OH, 0.65g, 2.62mmol), 6h is stirred at room temperature in DCC (0.59g, 2.86mmol) and DMAP (58mg, 0.48mmol)
To complete, it is filtered to remove white solid, acquired solution is concentrated under reduced pressure, compound 13 is then purified to obtain by silica gel column chromatography
(1.3g, 86.7%).Compound 13 (1.3g, 2.07mmol) is dissolved in DCM (7mL), is added TFA (3mL), 2h is stirred at room temperature extremely
Fully reacting is concentrated under reduced pressure and removes TFA and DCM, and saturated sodium bicarbonate aqueous solution (30mL) is added in residue, DCM (3 ×
It 30mL) extracts, merges organic phase, after saturated brine washs, successively through anhydrous Na2SO4It dries, filters, is concentrated under reduced pressure, finally
Compound 14 (0.80g, 73.4%) is purified to obtain by silica gel column chromatography.Compound 14 (0.80g, 1.51mmol) is dissolved in methanol
(MeOH, 10mL) is added Pd/C (containing Pd10%, 50mg), pricks hydrogen balloon, carry out gas exchanges three times, react at room temperature overnight extremely
Completely, gained reaction solution is filtered, compound 15 (700mg) is concentrated under reduced pressure to obtain；Compound 15 (280mg, 0.64mmol) is molten
In anhydrous THF (6mL), ice bath is added TEA (125 μ L, 0.90mmol), dropwise addition allyl chlorocarbonate (Alloc-Cl, 43 μ L,
0.70mmol), ice bath reacts 6h to complete, is then concentrated under reduced pressure and removes THF, and 10mL water is added, and adjusts pH to 2- with 1M HCl
3, after being extracted using DCM, merges organic phase, then successively washed with saturated brine, anhydrous Na2SO4It dries, filters, is concentrated under reduced pressure
Afterwards, compound 16 (170mg, 51%) is purified to obtain using silica gel column chromatography
MS(ESI)calcd.for C25H35N2O10 +[M+H]+=523.23, found 523.23.1H NMR(400MHz,
CDCl3) δ 7.33-7.18 (m, 5H), 6.16 (d, J=8.7Hz, 1H), 5.99-5.86 (m, 1H), 5.69 (d, J=4.2Hz,
1H), 5.37-5.20 (m, 3H), 4.86 (dd, J=9.3,2.2Hz, 1H), 4.69 (d, J=4.9Hz, 1H), 4.63 (d, J=
5.5Hz, 2H), 3.92 (dd, J=12.0,6.1Hz, 1H), 3.26 (s, 3H), 3.25-3.20 (m, 2H), 3.06 (s, 3H),
2.12 (s, 3H), 1.22 (d, J=6.4Hz, 3H), 1.16 (d, J=6.3Hz, 3H).13C NMR (100MHz, CDCl3) δ
171.14,169.88,167.93,167.43,156.89,134.98,131.48,128.49,127.44,125.98,116.52,
75.48,73.55,70.15,65.80,61.59,56.01,54.77,35.92,31.60,21.80,15.78,13.86。
The preparation of compound 25
By (2S, 3R)-(+) -2- amino -3- hydroxy-4-methyl valeric acid (3.01g, 20.4mmol) and p-methyl benzenesulfonic acid
(TsOH, 3.86g, 22.4mmol) is dissolved in toluene, is added benzyl alcohol (BnOH, 10mL), and 120 DEG C of back flow reactions are overnight to having reacted
Entirely, it is then concentrated under reduced pressure and removes toluene, saturated sodium bicarbonate aqueous solution is added in gained reaction solution, then (repeat three by DCM
Secondary, each 30mL) extraction, merge organic phase, is then washed with saturated brine and anhydrous Na2SO4It dries, filters, is concentrated under reduced pressure
Afterwards, silica gel column chromatography purifies to obtain compound 17 (3.46g, 71%).Compound 17 (1.90g, 8.0mmol) is dissolved in DCM
Triethylamine (2.0mL, 9.6mmol) and di-tert-butyl dicarbonate [(Boc) is added in (20mL)2O, 1.33mL, 8.8mmol], instead
It answers liquid chamber temperature to be stirred to react 4h to complete, is concentrated under reduced pressure and removes excess of triethylamine and DCM, chemical combination is then obtained by silica gel column chromatography
Object 18 (2.57g, 95.2%).Compound 17 (2.21g, 9.3mmol) is separately dissolved in the mixed solution (MeCN/ of acetonitrile and water
H2O, 20mL, v/v=1:1), NaHCO is added3(1.56g, 18.6mmol) and fluorenes methoxy carbonyl acyl succinimide (FmocOSu,
2.44g, 10.2mmol), 3.5h is stirred at room temperature to complete, is then filtered to remove white solid, then remove MeCN through being concentrated under reduced pressure,
Then the water phase in acquired solution is extracted with ethyl acetate (in triplicate, each 20mL), organic phase anhydrous Na2SO4It is dry
It is dry, it is concentrated under reduced pressure after filtering, then obtain compound 19 (2.04g, 48.1%) with silica gel column chromatography；By compound 19 (1.7g,
It 3.7mmol) is dissolved in DCM (15mL), (Boc) is added in ice bath2O (0.94mL, 4.1mmol) and DMAP (45mg,
0.37mmol), reaction 5h is stirred at room temperature in reaction solution, is concentrated under reduced pressure and removes DCM, and silica gel column chromatography purifies up to compound 20
(1.4g, 67.6%)；Compound 20 (1.35g, 2.42mmol) is dissolved in MeOH (10mL) again, addition Pt/C (contain Pt10%,
150mg), hydrogen balloon is pricked, gas exchanges three times are carried out, 18h is reacted at room temperature, is concentrated under reduced pressure after filtering, is purified with silica gel column chromatography
It obtains compound 21 (1.03g, 3 step yields 91.2%).By compound 18 (312mg, 1.0mmol) and compound 21 (576mg,
It 1.2mmol) is dissolved in DCM (6mL), DCC (250mg, 1.2mmol) and DMAP (23mg, 0.3mmol) is added, 3h is stirred at room temperature,
It is filtered to remove white solid, is concentrated under reduced pressure, silica gel column chromatography purifies to obtain compound 22 (544mg, 74.6%).Again by compound 22
(1.53g, 2.0mmol) is dissolved in DCM (10mL), is added diethylamine (4mL), and reaction 4h is stirred at room temperature, is concentrated under reduced pressure, passes through silica gel
Column chromatographs to obtain compound 23 (660mg, 58.0%).Compound 23 (510mg, 0.95mmol) is dissolved in DCM (5mL), ice bath adds
Enter triethylamine (185 μ L, 1.33mmol), then chloroacetic chloride (AcCl, 186 μ L, 1.70mmol) are added dropwise, ice bath reacts 0.5h, moves to
Room temperature reacts 1h to complete again, and reaction solution is washed with saturated brine, organic phase anhydrous Na2SO4After being concentrated under reduced pressure after drying, filtering,
Compound 24 (510mg, 96.1%) is obtained by silica gel column chromatography.Compound 24 (1.03g, 1.7mmol) is dissolved in MeOH again
In (10mL), Pd/C (containing Pd 10%, 150mg) is added, pricks hydrogen balloon and carries out gas exchanges three times, reacts at room temperature filtering for 24 hours
After be concentrated under reduced pressure, then purified with silica gel column chromatography and can obtain target compound white foam solid fragment compound 25
(740mg, yield 82.0%)
MS(ESI)calcd.for C24H41N2O10 +[M+H]+519.30 found 519.30.1H NMR(500MHz,
CDCl3) δ 6.36 (d, J=9.4Hz, 1H), 5.29 (d, J=9.6Hz, 1H), 5.16 (d, J=9.1Hz, 1H), 4.89 (d, J=
7.3Hz, 2H), 4.65 (d, J=9.9Hz, 1H), 4.35 (s, 3H), 2.07 (s, 3H), 2.02-1.95 (m, 1H), 1.92-1.87
(m, 1H), 1.47 (s, 9H), 1.46 (s, 9H), 1.00 (dd, J=13.6,6.5Hz, 6H), 0.95 (d, J=6.5Hz, 3H),
0.90 (d, J=6.6Hz, 3H).13C NMR(100MHz,CDCl3)δ172.64,171.83,169.44,155.83,152.82,
82.94,81.01,80.19,80.14,54.54,52.96,29.94,29.30,28.29,27.60,22.67,18.80,
18.68,18.17。
I -1 synthesis in solid state
2- chlorine trityl chloride (2-CTC, 397mg, 0.51mmol/g) resin is swollen 15min with DCM (4mL), filters off
DCM, addition Fmoc-N-Me-Ala-OH (260mg, 0.80mmol) and n,N-diisopropylethylamine (DIEA, 206mg,
1.60mmol) and DCM (3mL), room temperature reaction 2h filter off reaction solution, successively use DCM/MeOH/DIEA (4mL, 17:2:1, v/
V), DCM (4mL), DMF (4mL) and DCM (4mL) respectively washing 3 times.Piperidines/DMF solution (4mL) of 20 (v/v) % is added, instead
10min is answered, reaction solution is filtered off, three times using DMF (4mL) washing, the above purification process is repeated once to obtain compound (A1).Again
By Fmoc-Ala-OH (249mg, 0.80mmol), 2- (7- aoxidizes benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphoric acid
After salt (HATU, 304mg, 0.80mmol) and trimethylpyridine (collidine, 195mg, 1.60mmol) are dissolved in DMF (4mL),
Compound (A1) is added and reacts 2h, filters off reaction solution, DMF (4mL) is washed three times, and the piperidines/DMF for adding 20 (v/v) % is molten
Liquid (4mL) reacts 10min, filters off reaction solution, and DMF (4mL) is washed three times, and the above purification process is repeated once to obtain compound
(B1).Again by compound 6 (385mg, 0.80mmol), HATU (304mg, 0.80mmol) and collidine (195mg,
It 1.60mmol) being dissolved in DMF (4mL), reaction in compound (B1) is added and for 24 hours, filters off reaction solution, DMF (4mL) is washed three times,
Piperidines/DMF solution (4mL) of 20 (v/v) % is added, 10min is reacted, filters off reaction solution, DMF (4mL) is washed three times, above
Operation is repeated once.Again by compound 16 (418mg, 0.80mmol), HATU (304mg, 0.80mmol) and collidine
(195mg, 1.60mmol) is dissolved in DMF (4mL), and reaction in compound (C1) is added and for 24 hours, filters off reaction solution, DMF (4mL) is washed
It washs three times, adds DCM (3mL), Ar brushes lower addition phenylsilane (PhSiH3, 216mg, 2.0mmol) and Pd (PPh3)4
(23mg, 0.02mmol), Ar brush lower reaction 30min, filter off reaction solution, successively use DCM (4mL), DMF (4mL) and DCM
(4mL) respectively washing 3 times.Again by compound 25 (156mg, 0.30mmol), HATU (115mg, 0.30mmol) and collidine
(73mg, 0.60mmol) is dissolved in DMF (2mL), and 35 DEG C of reactions in compound (D1) are added and for 24 hours, filter off reaction solution, DMF (4mL)
Three times, the above operation is repeated once to obtain compound (E1) for washing.To addition trifluoroacetic acid/triisopropyl silicon in compound (E1)
Alkane/methylene chloride TFA/TIPS/DCM (4mL, 19/0.5/0.5, v/v) reacts at room temperature 1h, filters and collect filtrate, will precipitate
Object respectively washes twice to obtain cleaning solution, merging filtrate and cleaning solution with TFA (2mL) and DCM (2mL) again, and vacuum distillation is concentrated to give
Crude product purifies to obtain I -1 chain precursor of cyclic ester peptide. i.e. compound (F1) 4.0mg by preparative HPLC.Finally by compound
(F1) (4.0mg, 0.0035mmol) is dissolved in anhydrous DMF (4mL), and DIEA (1.4mg, 0.0105mmol) and hexafluorophosphoric acid is added
Benzotriazole -1- base-oxygroup tripyrrole alkyl phosphorus PyBop (1.8mg, 0.0035mmol) reacts 2h to complete at room temperature, leads to
It crosses preparative HPLC and purifies to obtain cyclic ester peptide. I -1 (0.06mg, white powdery solid).
MS(ESI)calcd.for C52H76N7O15Se+[M+H]+1118.46, found 1118.46.1H NMR
(600MHz,CDCl3) δ 8.56 (d, J=8.8Hz, 1H), 8.19-8.14 (m, 2H), 7.86 (d, J=7.0Hz, 1H), 7.79
(d, J=8.8Hz, 1H), 7.36-7.28 (m, 9H), 7.14 (dd, J=7.1,2.4Hz, 2H), 5.69 (d, J=1.9Hz, 1H),
5.35 (d, J=9.3Hz, 1H), 5.30 (dd, J=6.4,1.9Hz, 1H), 5.13 (d, J=8.0Hz, 1H), 5.03 (dd, J=
9.8,3.8Hz, 2H), 4.99 (d, J=10.3Hz, 1H), 4.72 (dd, J=8.6,6.9Hz, 1H), 4.39 (d, J=10.0Hz,
1H), 3.88 (s, 1H), 3.81 (d, J=9.2Hz, 2H), 3.72 (dd, J=10.1,5.9Hz, 1H), 3.42 (s, 3H), 3.22
(dd, J=13.2,9.8Hz, 1H), 3.01 (s, 3H), 2.90 (dd, J=13.2,5.2Hz, 1H), 2.87 (s, 3H), 2.64 (s,
3H), 2.23 (d, J=4.6Hz, 2H), 2.21 (s, 3H), 2.12 (s, 3H), 1.94-1.81 (m, 4H), 1.75 (d, J=
7.5Hz, 3H), 1.44 (d, J=7.1Hz, 3H), 1.34 (d, J=6.3Hz, 3H), 1.08 (d, J=6.7Hz, 3H), 1.02 (d,
J=6.7Hz, 3H), 0.99 (d, J=6.8Hz, 3H), 0.90 (d, J=6.8Hz, 3H)13C NMR(150MHz,CDCl3)δ
175.1,174.9,173.0,171.6,170.7,170.5,168.2,168.0,166.4,163.8,141.1,134.8,
132.9,130.5,129.9,128.9,128.7,128.6,128.4,127.7,123.1,79.5,76.9,74.7,71.8,
71.1,65.4,57.1,55.6,55.2,55.0,48.1,46.2,40.1,38.9,37.2,33.8,30.7,30.0,29.1,
29.0,22.6,22.5,20.0,19.0,18.8,18.3,16.6,16.5,16.4,16.3,14.5,14.2.
The synthesis of embodiment 2:I-2
The present embodiment difference from example 1 is that, compound 1 be N- fluorenylmethyloxycarbonyl-N- methyl-L- Kosé ammonia
Sour (2.01g, 5mmol).
It obtains cyclic ester peptide. I-2 (0.10mg, white powdery solid).MS(ESI)calcd.for C53H78N7O15Se+[M+H]+
1132.47, found 1132.47.1H NMR(600MHz,CDCl3) δ 8.57 (d, J=8.8Hz, 1H), 8.20-8.10 (m,
2H), 7.86 (d, J=7.0Hz, 1H), 7.77 (d, J=8.8Hz, 1H), 7.34-7.24 (m, 9H), 7.13 (dd, J=7.1,
2.4Hz, 2H), 5.69 (d, J=1.9Hz, 1H), 5.35 (d, J=9.3Hz, 1H), 5.31 (dd, J=6.4,1.9Hz, 1H),
5.13 (d, J=8.0Hz, 1H), 5.03 (dd, J=9.8,3.8Hz, 2H), 4.98 (d, J=10.3Hz, 1H), 4.72 (dd, J=
8.6,6.9Hz, 1H), 4.39 (d, J=10.0Hz, 1H), 3.89 (s, 1H), 3.81 (d, J=9.2Hz, 2H), 3.73 (dd, J=
10.1,5.9Hz, 1H), 3.42 (s, 3H), 3.22 (dd, J=13.2,9.8Hz, 1H), 3.00 (s, 3H), 2.90 (dd, J=
13.2,5.2Hz, 1H), 2.89 (s, 3H), 2.64 (s, 3H), 2.24 (d, J=4.6Hz, 2H), 2.21 (s, 3H), 2.12 (s,
3H), 1.94-1.81 (m, 6H), 1.75 (d, J=7.5Hz, 3H), 1.44 (d, J=7.1Hz, 3H), 1.33 (d, J=6.3Hz,
3H), 1.08 (d, J=6.7Hz, 3H), 1.02 (d, J=6.7Hz, 3H), 0.99 (d, J=6.8Hz, 3H), 0.91 (d, J=
6.8Hz,3H).13C NMR(150MHz,CDCl3)δ175.1,174.9,173.1,171.8,170.7,170.5,168.2,
168.1,166.4,163.8,141.2,134.8,132.8,130.5,129.9,128.9,128.7,128.7,128.4,
127.7,123.1,79.4,76.9,74.8,71.8,71.1,65.4,57.1,55.7,55.2,55.0,48.2,46.2,40.1,
38.9,37.3,33.8,30.7,30.0,29.1,29.0,24.5,22.6,22.4,20.0,19.0,18.8,18.3,16.7,
16.5,16.4,16.3,14.4,14.2.
The synthesis of embodiment 3:I-3
The present embodiment difference from example 1 is that, compound 6 be N- fluorenylmethyloxycarbonyl-N- methyl-O- benzyl-
Serine (356mg, 0.8mmol).
It obtains cyclic ester peptide. I-3 (0.09mg, white powdery solid).MS(ESI)calcd.for C53H78N7O16 +[M+H]+
1068.55, found 1068.55.1H NMR(600MHz,CDCl3) δ 8.56 (d, J=8.8Hz, 1H), 8.19-8.14 (m,
2H), 7.87 (d, J=7.0Hz, 1H), 7.78 (d, J=8.8Hz, 1H), 7.36-7.28 (m, 9H), 7.18-7.07 (m, 2H),
5.69 (d, J=1.9Hz, 1H), 5.35 (d, J=9.3Hz, 1H), 5.33-5.26 (m, 1H), 5.14 (d, J=8.0Hz, 1H),
5.07-4.96 (m, 3H), 4.72 (dd, J=8.6,6.9Hz, 1H), 4.61 (d, 2H), 4.39 (d, J=10.0Hz, 1H), 3.88
(s, 1H), 3.81 (d, J=9.2Hz, 2H), 3.74-3.68 (m, 1H), 3.42 (s, 3H), 3.40-3.22 (m, 3H), 3.02 (s,
3H), 2.90 (dd, J=13.2,5.2Hz, 1H), 2.88 (s, 3H), 2.64 (s, 3H), 2.23 (d, J=4.6Hz, 2H), 2.21
(s, 3H), 2.12 (s, 3H), 1.94-1.80 (m, 2H), 1.75 (d, J=7.5Hz, 3H), 1.44 (d, J=7.1Hz, 3H),
1.34 (d, J=6.3Hz, 3H), 1.08 (d, J=6.7Hz, 3H), 1.04-0.95 (m, 6H), 0.90 (d, J=6.8Hz, 3H)
.13C NMR(150MHz,CDCl3)δ175.1,174.8,173.0,171.6,170.6,170.5,168.2,168.1,166.4,
163.8,141.1,134.8,132.9,130.5,129.9,128.9,128.7,128.6,128.4,127.7,123.1,79.5,
76.8,74.7,72.2,71.8,71.0,67.3,65.4,57.1,55.6,55.3,55.0,48.1,46.2,40.2,38.9,
37.2,33.8,30.7,30.0,29.1,22.6,22.5,20.0,19.0,18.8,18.3,16.6,16.5,16.4,16.3,
14.5,14.2.
Embodiment 4: active testing
(1) cell culture: stablize expression M1Chinese hamster ovary (CHO) cell CHO-k1 (CHO- of muscarinic receptor
M1) buy from American Type Culture collection warehousing ATCC, CHO-M1Cell is with added with 10% (v/v) fetal calf serum (FBS), 1%
(v/v) mycillin is dual anti-and Ham ' the s F12 culture medium of 0.25mg/mL G418 is cultivated.All cells maintain 37
DEG C wet environment in (95% air and 5% oxygen).
(2) 1 gained chemical compounds I -1 of embodiment is configured to the mother liquor of 20mM, with analysis buffer (20mM hydroxyethyl piperazine
Ethanesulfonic acid, pH 7.4,1mM CaCl2,1mM MgCl2) it is diluted to 50nM, 384 orifice plates of parallel three parts of additions (1 hole μ L/).
(3) inositol monophosphate (IP1) analysis: use non-enzymatic cell dissociation buffer by CHO-M1Cell is dissociated from culture dish
Get off, is then resuspended in analysis buffer shown in (2) the step of added with 0.2% bovine serum albumin(BSA) (BSA), keeps cell close
Degree contains 2,000,000 cells for every milliliter.Chemical compounds I -1 and cell (every Kong Yiwan cell) are incubated for 37 DEG C in 384 orifice plates
Educate 1h.After the completion of incubation, carbachol is dissolved in analysis liquid and 20mM LiCl is added.Again by carbachol with ultimate density be 3
μM concentration be added in every hole, in 37 DEG C of incubations 1h and be incubated at room temperature 15min.Reuse IP1-Gq-kit (Cisbio) reagent
The content of box detection IP1.
(4) pass through measurement IP1Amount evaluate it to Gq protein inhibiting activity.
The results show that cyclic ester peptide. I -1 inhibits the IC of Gq albumen50It is 0.1-1 μM, cyclic ester peptide. I -2 and I -3 inhibits Gq albumen
IC50It is 1-10 μM.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent substitute mode, be included within the scope of the present invention.
Claims (7)
1. a kind of cyclic ester peptides Gq protein inhibitor, which is characterized in that the inhibitor be such as logical formula (I) compound represented or
Its pharmaceutically acceptable salt or ester or solvate:
Wherein,
N is selected from 0,1,2；
Y is selected from O, S, Se ,-CH2,-NH- ,-CONH- ,-NHCO- ,-COO- or-OOC-；
Z is selected from benzyl, aryl, heteroaryl, naphthenic base, Heterocyclylalkyl, straight chained alkyl, guanidine radicals, phosphate, naphthalene, quinolyl, benzene
And furyl or indyl.
2. cyclic ester peptides Gq protein inhibitor according to claim 1, it is characterised in that:
In the Z, heteroaryl and Heterocyclylalkyl are respectively containing 1~3 hetero atom selected from N, O or S.
3. cyclic ester peptides Gq protein inhibitor according to claim 1, it is characterised in that:
The Z is replaced by 1~3 identical or different R；
R is hydrogen, fluorine, chlorine, smell, at least one of iodine, hydroxyl, nitro, cyano, trifluoromethyl, methoxyl group and trifluoromethoxy.
4. cyclic ester peptides Gq protein inhibitor according to claim 1, it is characterised in that: change shown in the logical formula (I)
Closing object has structure shown in I-1~I-33:
5. a kind of method for preparing any one of Claims 1 to 4 cyclic ester peptides Gq protein inhibitor, which is characterized in that institute
State method specifically includes the following steps:
(1) by 2- chlorine trityl chloride resin, N- fluorenes methoxy carbonyl acyl group-N- methyl-L-alanine and N, N- diisopropylethylamine
It is reacted after being mixed in DCM, adds piperidines/DMF mixed solution after the reaction was completed the reaction was continued and obtain compound (A)；
(2) by N- fluorenes methoxy carbonyl acyl group-N- methyl-L-alanine, 2- (7- aoxidizes benzotriazole)-N, N, N', N'- tetramethyl
Urea hexafluorophosphate, trimethylpyridine are dissolved in after DMF to be reacted with after compound (A) obtained by step (1), reaction complete again plus
Enter piperidines/DMF mixed solution the reaction was continued to obtain compound (B)；
(3) by compound G, 2- (7- aoxidize benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphate and trimethyl pyrrole
Pyridine be dissolved in after DMF with compound (B) hybrid reaction obtained by step (2), add piperidines, the reaction was continued obtains compound (C)；Its
Middle compound G general structure is as follows:
Compound G
Wherein Z, Y, n all have meaning described in claim 1；
(4) by compound 16,2- (7- aoxidizes benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphate and trimethyl pyrrole
Pyridine is dissolved in after DMF and compound (C) hybrid reaction obtained by step (3) is added, and then adds phenylsilane, Pd (PPh3)4And DCM
The reaction was continued, obtains compound (D)；Wherein, the general structure of compound 16 is as follows:
(5) by compound 25,2- (7- aoxidizes benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphate and trimethyl pyrrole
Pyridine is dissolved in DMF, then with compound (D) hybrid reaction obtained by step (4), obtains compound (E), wherein the structure of compound 25
Formula is as follows:
(6) it by after compound (E) obtained by step (5) and trifluoroacetic acid/tri isopropyl silane/methylene chloride hybrid reaction, filters
And filtrate is collected, filtrate is concentrated to get compound (F), i.e. cyclic ester peptide. chain precursor；
(7) after compound (F) obtained by step (6) being dissolved in DMF, with n,N-diisopropylethylamine and hexafluorophosphoric acid benzotriazole-
1- base-oxygroup tripyrrole alkyl phosphorus hybrid reaction, obtains the cyclic ester peptides Gq protein inhibitor as shown in formula (I).
6. any one cyclic ester peptides Gq protein inhibitor is preparing antitumor, antithrombotic and anti-heavy breathing according to claim 1~4
It is applied in asthma drug.
7. a kind of Pharmaceutical composition comprising described in any item cyclic ester peptides Gq protein inhibitors of Claims 1 to 4 and at least
A kind of pharmaceutic adjuvant.
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Citations (2)
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| CN104684546A (en) * | 2012-06-07 | 2015-06-03 | 哈佛大学校长及研究员协会 | Nanotherapeutics for drug targeting |
| WO2019060781A1 (en) * | 2017-09-22 | 2019-03-28 | Washington University | Targeted pharmacological therapeutics in uveal melanoma |
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| CN104684546A (en) * | 2012-06-07 | 2015-06-03 | 哈佛大学校长及研究员协会 | Nanotherapeutics for drug targeting |
| WO2019060781A1 (en) * | 2017-09-22 | 2019-03-28 | Washington University | Targeted pharmacological therapeutics in uveal melanoma |
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| XIAO-FENG XIONG: "Total synthesis and structure–activity relationship studies of a series of selective G protein inhibitors", 《NATURE CHEMISTRY》 * |
| ZHANG HANG 等: "Structure-Activity Relationship Studies of the Cyclic Depsipeptide Natural Product YM-254890, Targeting the Gq Protein", 《CHEMMEDCHEM》 * |
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