Amylin receptor AMY1/3

Amylin is an important but poorly understood 37-amino acid glucose-regulating hormone with considerable potential to cure metabolic disorders. Amylin is a member of the calcitonin (CT) peptide family, which comprises CT itself, CGRP having two forms (αCGRP and βCGRP), adrenomedullin (AM), and AM2 (intermedin). Amylin is a key neuroendocrine hormone produced with insulin in pancreatic beta cells. Amylin maintains glucose homeostasis by decreasing gastric emptying, reducing the release of the counterregulatory hormone glucagon, and producing postprandial satiety. Amylin, a glucose-regulating and satiety-inducing hormone, reduces postprandial blood sugar increases and overeating. [1]

Amylin is an important, but poorly understood, 37 amino acid glucoregulatory hormone with great potential to target metabolic diseases. A working example that the amylin system is one worth developing is the FDA-approved drug used in insulin-requiring diabetic patients, pramlintide. However, certain characteristics of pramlintide pharmacokinetics and formulation leave considerable room for further development of amylin-mimetic compounds. Given that amylin-mimetic drug design and development is an active area of research, surprisingly little is known about the structure/function relationships of amylin. This is largely due to the unfavourable aggregative and solubility properties of the native peptide sequence, which are further complicated by the composition of amylin receptors. These are complexes of the calcitonin receptor with receptor activity-modifying proteins. This review explores what is known of the structure-function relationships of amylin and provides insights that can be drawn from the closely related peptide, CGRP. We also describe how this information is aiding the development of more potent and stable amylin mimetics, including peptide hybrids. [1]

[1]. Amylin structure-function relationships and receptor pharmacology: implications for Amylin mimetic drug development. Br J Pharmacol. 2016 Jun;173(12):1883-98.

From available data thus far, it is becoming increasingly apparent that whilst the two‐domain model of binding and activation for family B receptor peptide ligands is useful, this is only to a limited degree. N‐terminal ring fragments retain biological activity (Rossowski et al., 1997), and C‐terminal fragments are often antagonists and retain binding to receptors (Barwell et al., 2012) validating these facets of the model; however, the data are not always so cleanly defined. Also questionable is the degree of importance of the disulphide ‘activation loop’ and C‐terminal tyrosine amide in the amylin peptide. The studies for amylin investigating their roles have not been carried out in biological assays with defined amylin receptors or in measuring canonical amylin‐mediated physiological actions (Roberts et al., 1989). With CGRP, breaking the N‐terminal disulphide still resulted in partial agonists in some biological assays. There are suitable suggestions as to the secondary structure the native amylin peptide adopts although, in solution, it is likely to be disordered and capriciously change structure (He et al., 2015). Information from CD and NMR studies utilize detergent membranes to mimic cellular membrane/peptide interactions, which are not ideal as they naturally instigate helical conformations, and these vary depending on solvent used and/or micellar composition (Watkins et al., 2012). In order to be certain, crystal structures of amylin or pramlintide bound to an amylin receptor are needed. The crystal structures for a CGRP27–37 analogue and AM37–52 offer useful insights (Booe et al., 2015) but fragments only tell part of the story, and N‐terminal interactions with receptor juxtamembrane regions are excluded in these models. The scope for peptide modification strategies to develop new amylin mimetics is substantial. However, to drive drug design and development, more information is needed to understand amylin and how it acts to elicit physiological responses and, thus, how its structure influences function. Metabolism and glucoregulation are enormously complex physiological processes requiring multifaceted hormonal and enzymic responses. In the future, it is likely to be combination therapies that will be the most useful to effectively target diseases such as diabetes and obesity.[1]

C167H272N52O53S2

3920.39321422577

3978.987

124447-81-0

163339198

H-Lys-Cys(1)-Asn-Thr-Ala-Thr-Cys(1)-Ala-Thr-Gln-Arg-Leu-Ala-Asn-Phe-Leu-Val-Arg-Ser-Ser-Asn-Asn-Leu-Gly-Pro-Val-Leu-Pro-Pro-Thr-Asn-Val-Gly-Ser-Asn-Thr-Tyr-NH2; Lys-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Asn-Phe-Leu-Val-Arg-Ser-Ser-Asn-Asn-Leu-Gly-Pro-Val-Leu-Pro-Pro-Thr-Asn-Val-Gly-Ser-Asn-Thr-Tyr-NH2 (Disulfide bridge: Cys2-Cys7); L-lysyl-L-cysteinyl-L-asparagyl-L-threonyl-L-alanyl-L-threonyl-L-cysteinyl-L-alanyl-L-threonyl-L-glutaminyl-L-arginyl-L-leucyl-L-alanyl-L-asparagyl-L-phenylalanyl-L-leucyl-L-valyl-L-arginyl-L-seryl-L-seryl-L-asparagyl-L-asparagyl-L-leucyl-glycyl-L-prolyl-L-valyl-L-leucyl-L-prolyl-L-prolyl-L-threonyl-L-asparagyl-L-valyl-glycyl-L-seryl-L-asparagyl-L-threonyl-L-tyrosinamide (2->7)-disulfide

KCNTATCATQRLANFLVRSSNNLGPVLPPTNVGSNTY-NH2 (Disulfide bridge: Cys2-Cys7); KCNTATCATQRLANFLVRSSNNLGPVLPPTNVGSNTY

White to off-white solid powder

59

61

112

278

9610

37

S1C[C@@H](C(N[C@@H](C)C(N[C@@H]([C@@H](C)O)C(N[C@@H](CCC(N)=O)C(N[C@H](C(N[C@H](C(N[C@@H](C)C(N[C@@H](CC(N)=O)C(N[C@@H](CC2C=CC=CC=2)C(N[C@H](C(N[C@@H](C(C)C)C(N[C@@H](CCCNC(=N)N)C(N[C@@H](CO)C(N[C@@H](CO)C(N[C@@H](CC(N)=O)C(N[C@@H](CC(N)=O)C(N[C@@H](CC(C)C)C(NCC(N2CCC[C@H]2C(N[C@@H](C(C)C)C(N[C@@H](CC(C)C)C(N2CCC[C@H]2C(N2CCC[C@H]2C(N[C@H](C(N[C@H](C(N[C@H](C(NCC(N[C@H](C(N[C@H](C(N[C@H](C(N[C@H](C(N)=O)CC2C=CC(=CC=2)O)=O)[C@@H](C)O)=O)CC(N)=O)=O)CO)=O)=O)C(C)C)=O)CC(N)=O)=O)[C@@H](C)O)=O)=O)=O)=O)=O)=O)=O)=O)=O)=O)=O)=O)=O)=O)CC(C)C)=O)=O)=O)=O)CC(C)C)=O)CCCNC(=N)N)=O)=O)=O)=O)NC([C@H]([C@@H](C)O)NC([C@H](C)NC([C@H]([C@@H](C)O)NC([C@H](CC(N)=O)NC([C@H](CS1)NC([C@H](CCCCN)N)=O)=O)=O)=O)=O)=O

FEOIOOASAUJGKY-HDCGBPIBSA-N

InChI=1S/C167H272N52O53S2.C2H4O2/c1-72(2)53-95(136(243)185-66-122(237)217-50-30-38-111(217)154(261)211-125(78(13)14)158(265)204-105(56-75(7)8)164(271)219-52-32-40-113(219)165(272)218-51-31-39-112(218)155(262)216-130(86(22)227)162(269)203-104(64-120(176)235)146(253)209-123(76(9)10)156(263)184-65-121(236)189-106(67-220)149(256)201-102(62-118(174)233)147(254)215-129(85(21)226)161(268)193-94(131(177)238)57-88-41-43-89(228)44-42-88)195-143(250)100(60-116(172)231)199-144(251)101(61-117(173)232)200-150(257)107(68-221)206-151(258)108(69-222)205-138(245)92(37-29-49-183-167(180)181)191-157(264)124(77(11)12)210-145(252)97(55-74(5)6)197-141(248)98(58-87-33-24-23-25-34-87)198-142(249)99(59-115(171)230)194-132(239)79(15)186-140(247)96(54-73(3)4)196-137(244)91(36-28-48-182-166(178)179)190-139(246)93(45-46-114(170)229)192-160(267)127(83(19)224)212-133(240)80(16)187-152(259)109-70-273-274-71-110(207-135(242)90(169)35-26-27-47-168)153(260)202-103(63-119(175)234)148(255)214-126(82(18)223)159(266)188-81(17)134(241)213-128(84(20)225)163(270)208-109;1-2(3)4/h23-25,33-34,41-44,72-86,90-113,123-130,220-228H,26-32,35-40,45-71,168-169H2,1-22H3,(H2,170,229)(H2,171,230)(H2,172,231)(H2,173,232)(H2,174,233)(H2,175,234)(H2,176,235)(H2,177,238)(H,184,263)(H,185,243)(H,186,247)(H,187,259)(H,188,266)(H,189,236)(H,190,246)(H,191,264)(H,192,267)(H,193,268)(H,194,239)(H,195,250)(H,196,244)(H,197,248)(H,198,249)(H,199,251)(H,200,257)(H,201,256)(H,202,260)(H,203,269)(H,204,265)(H,205,245)(H,206,258)(H,207,242)(H,208,270)(H,209,253)(H,210,252)(H,211,261)(H,212,240)(H,213,241)(H,214,255)(H,215,254)(H,216,262)(H4,178,179,182)(H4,180,181,183);1H3,(H,3,4)/t79-,80-,81-,82+,83+,84+,85+,86+,90-,91-,92-,93-,94-,95-,96-,97-,98-,99-,100-,101-,102-,103?,104-,105-,106-,107-,108-,109-,110?,111-,112-,113-,123-,124-,125-,126-,127-,128?,129-,130-;/m0./s1

acetic acid;(2S)-N-[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-4-amino-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-4-amino-1-[[(2S)-4-amino-1-[[(2S)-1-[[2-[(2S)-2-[[(2S)-1-[[(2S)-1-[(2S)-2-[(2S)-2-[[(2S,3R)-1-[[(2S)-4-amino-1-[[(2S)-1-[[2-[[(2S)-1-[[(2S)-4-amino-1-[[(2S,3R)-1-[[(2S)-1-amino-3-(4-hydroxyphenyl)-1-oxopropan-2-yl]amino]-3-hydroxy-1-oxobutan-2-yl]amino]-1,4-dioxobutan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-2-oxoethyl]amino]-3-methyl-1-oxobutan-2-yl]amino]-1,4-dioxobutan-2-yl]amino]-3-hydroxy-1-oxobutan-2-yl]carbamoyl]pyrrolidine-1-carbonyl]pyrrolidin-1-yl]-4-methyl-1-oxopentan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]carbamoyl]pyrrolidin-1-yl]-2-oxoethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-1,4-dioxobutan-2-yl]amino]-1,4-dioxobutan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-5-carbamimidamido-1-oxopentan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-1-oxo-3-phenylpropan-2-yl]amino]-1,4-dioxobutan-2-yl]amino]-1-oxopropan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-carbamimidamido-1-oxopentan-2-yl]-2-[[(2S,3R)-2-[[(2S)-2-[[(4R,10S,13S)-16-(2-amino-2-oxoethyl)-19-[[(2S)-2,6-diaminohexanoyl]amino]-7,13-bis[(1R)-1-hydroxyethyl]-10-methyl-6,9,12,15,18-pentaoxo-1,2-dithia-5,8,11,14,17-pentazacycloicosane-4-carbonyl]amino]propanoyl]amino]-3-hydroxybutanoyl]amino]pentanediamide

Amylin (rat); 124447-81-0;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ~100 mg/mL (~25.51 mM)
DMSO : ~50 mg/mL (~12.75 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)