Need Assistance?

US & Canada:
+
UK: +

Our Highlights

GMP Grade Efficient workshop for GMP production.
Optimal Prices Buying products on this site guarantees the best price!
Commercial Batches Independent GMP manufacturing suites that handle commercial batches
Prompt Delivery Warehouses in multiple cities to ensure timely delivery
Flexible Quantity Quantities available from milligrams to ton

Difopein

* Please kindly note that our products are not to be used for therapeutic purposes and cannot be sold to patients.

Difopein has been found to be a 14.3.3 protein inhibitor and could induce apoptosis expressed in COS-7 and some cancer cells.

Category

Peptide Inhibitors

Catalog number

BAT-010334

CAS number

396834-58-5

Molecular Formula

C273H424N76O89S6

Molecular Weight

6387.17

Size	Price	Stock	Quantity
1 mg	$838	In stock
5 mg	$1999	In stock

Specification
Reference Reading

IUPAC Name

(2S)-2-[[(2S)-2-[[2-[[(2S)-2-[[(2S)-2-[[2-[[(2S)-1-[(2S)-2-[[(2R)-2-[[(2S)-2-[[(2S)-4-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-1-[(2S)-2-[[(2R)-2-[[(2S)-2-[[(2S)-1-[(2S)-2-[[2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[2-[[(2S)-2-[[(2S)-2-[[2-[[(2S)-1-[(2S)-2-[[(2R)-2-[[(2S)-2-[[(2S)-4-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-1-[(2S)-2-[[(2R)-2-[[(2S)-2-[[(2S)-1-[(2S)-2-[[2-[[(2S)-2-[[(2S)-2-[[(2S)-2-amino-3-hydroxypropanoyl]amino]propanoyl]amino]-3-carboxypropanoyl]amino]acetyl]amino]propanoyl]pyrrolidine-2-carbonyl]amino]-3-(1H-imidazol-4-yl)propanoyl]amino]-3-sulfanylpropanoyl]amino]-3-methylbutanoyl]pyrrolidine-2-carbonyl]amino]-5-carbamimidamidopentanoyl]amino]-3-carboxypropanoyl]amino]-4-methylpentanoyl]amino]-3-hydroxypropanoyl]amino]-3-(1H-indol-3-yl)propanoyl]amino]-4-methylpentanoyl]amino]-3-carboxypropanoyl]amino]-4-methylpentanoyl]amino]-4-carboxybutanoyl]amino]propanoyl]amino]-4-oxobutanoyl]amino]-4-methylsulfanylbutanoyl]amino]-3-sulfanylpropanoyl]amino]-4-methylpentanoyl]pyrrolidine-2-carbonyl]amino]acetyl]amino]propanoyl]amino]propanoyl]amino]acetyl]amino]-4-methylpentanoyl]amino]-3-carboxypropanoyl]amino]-3-hydroxypropanoyl]amino]propanoyl]amino]-3-carboxypropanoyl]amino]acetyl]amino]propanoyl]pyrrolidine-2-carbonyl]amino]-3-(1H-imidazol-4-yl)propanoyl]amino]-3-sulfanylpropanoyl]amino]-3-methylbutanoyl]pyrrolidine-2-carbonyl]amino]-5-carbamimidamidopentanoyl]amino]-3-carboxypropanoyl]amino]-4-methylpentanoyl]amino]-3-hydroxypropanoyl]amino]-3-(1H-indol-3-yl)propanoyl]amino]-4-methylpentanoyl]amino]-3-carboxypropanoyl]amino]-4-methylpentanoyl]amino]-4-carboxybutanoyl]amino]propanoyl]amino]-4-oxobutanoyl]amino]-4-methylsulfanylbutanoyl]amino]-3-sulfanylpropanoyl]amino]-4-methylpentanoyl]pyrrolidine-2-carbonyl]amino]acetyl]amino]propanoyl]amino]propanoyl]amino]acetyl]amino]-4-methylpentanoyl]amino]pentanedioic acid

Synonyms

Difopein; 396834-58-5; HB2038; AKOS024456954; C338H529N97O105S11; IVCHTTATSPISAVTCPPGENLCYRKMWCDAFCSSRGKVVELGCAATCPSKKPYEEVTCCSTDKCNPHPKQRPG

Appearance

White Lyophilized Solid

Purity

≥95% by HPLC

Sequence

SADGAPHCVPRDLSWLDLEANMCLPGAAGLDSADGAPHCVPRDLSWLDLEANMCLPGAAGLE

Storage

Store at -20°C

Solubility

Soluble in DMSO

InChI

InChI=1S/C273H424N76O89S6/c1-122(2)79-160(233(399)313-159(271(437)438)63-66-206(366)367)305-202(360)110-287-216(382)134(25)299-218(384)136(27)295-198(356)108-291-259(425)190-55-43-73-346(190)267(433)181(87-130(17)18)333-255(421)186(116-439)338-231(397)157(67-77-443-35)309-245(411)172(93-196(275)354)314-221(387)139(30)302-227(393)155(61-64-204(362)363)307-235(401)162(81-124(5)6)320-249(415)178(99-211(376)377)329-237(403)165(84-127(11)12)319-242(408)169(90-145-103-286-152-52-40-38-50-149(145)152)325-254(420)185(115-353)337-240(406)167(86-129(15)16)323-248(414)177(98-210(374)375)327-230(396)154(54-42-70-284-273(279)280)312-262(428)195-60-48-76-349(195)270(436)215(133(23)24)343-258(424)189(119-442)341-244(410)171(92-147-105-282-121-294-147)332-264(430)193-58-46-72-345(193)266(432)143(34)298-201(359)107-290-226(392)175(96-208(370)371)317-223(389)141(32)304-252(418)183(113-351)335-251(417)180(101-213(380)381)328-234(400)161(80-123(3)4)306-203(361)111-288-217(383)135(26)300-219(385)137(28)296-199(357)109-292-260(426)191-56-44-74-347(191)268(434)182(88-131(19)20)334-256(422)187(117-440)339-232(398)158(68-78-444-36)310-246(412)173(94-197(276)355)315-222(388)140(31)303-228(394)156(62-65-205(364)365)308-236(402)163(82-125(7)8)321-250(416)179(100-212(378)379)330-238(404)164(83-126(9)10)318-241(407)168(89-144-102-285-151-51-39-37-49-148(144)151)324-253(419)184(114-352)336-239(405)166(85-128(13)14)322-247(413)176(97-209(372)373)326-229(395)153(53-41-69-283-272(277)278)311-261(427)194-59-47-75-348(194)269(435)214(132(21)22)342-257(423)188(118-441)340-243(409)170(91-146-104-281-120-293-146)331-263(429)192-57-45-71-344(192)265(431)142(33)297-200(358)106-289-225(391)174(95-207(368)369)316-220(386)138(29)301-224(390)150(274)112-350/h37-40,49-52,102-105,120-143,150,153-195,214-215,285-286,350-353,439-442H,41-48,53-101,106-119,274H2,1-36H3,(H2,275,354)(H2,276,355)(H,281,293)(H,282,294)(H,287,382)(H,288,383)(H,289,391)(H,290,392)(H,291,425)(H,292,426)(H,295,356)(H,296,357)(H,297,358)(H,298,359)(H,299,384)(H,300,385)(H,301,390)(H,302,393)(H,303,394)(H,304,418)(H,305,360)(H,306,361)(H,307,401)(H,308,402)(H,309,411)(H,310,412)(H,311,427)(H,312,428)(H,313,399)(H,314,387)(H,315,388)(H,316,386)(H,317,389)(H,318,407)(H,319,408)(H,320,415)(H,321,416)(H,322,413)(H,323,414)(H,324,419)(H,325,420)(H,326,395)(H,327,396)(H,328,400)(H,329,403)(H,330,404)(H,331,429)(H,332,430)(H,333,421)(H,334,422)(H,335,417)(H,336,405)(H,337,406)(H,338,397)(H,339,398)(H,340,409)(H,341,410)(H,342,423)(H,343,424)(H,362,363)(H,364,365)(H,366,367)(H,368,369)(H,370,371)(H,372,373)(H,374,375)(H,376,377)(H,378,379)(H,380,381)(H,437,438)(H4,277,278,283)(H4,279,280,284)/t134-,135-,136-,137-,138-,139-,140-,141-,142-,143-,150-,153-,154-,155-,156-,157-,158-,159-,160-,161-,162-,163-,164-,165-,166-,167-,168-,169-,170-,171-,172-,173-,174-,175-,176-,177-,178-,179-,180-,181-,182-,183-,184-,185-,186-,187-,188-,189-,190-,191-,192-,193-,194-,195-,214-,215-/m0/s1

InChI Key

NTCJKZSYQTZRQE-CSMGIIAWSA-N

Canonical SMILES

CC(C)CC(C(=O)NC(CCC(=O)O)C(=O)O)NC(=O)CNC(=O)C(C)NC(=O)C(C)NC(=O)CNC(=O)C1CCCN1C(=O)C(CC(C)C)NC(=O)C(CS)NC(=O)C(CCSC)NC(=O)C(CC(=O)N)NC(=O)C(C)NC(=O)C(CCC(=O)O)NC(=O)C(CC(C)C)NC(=O)C(CC(=O)O)NC(=O)C(CC(C)C)NC(=O)C(CC2=CNC3=CC=CC=C32)NC(=O)C(CO)NC(=O)C(CC(C)C)NC(=O)C(CC(=O)O)NC(=O)C(CCCNC(=N)N)NC(=O)C4CCCN4C(=O)C(C(C)C)NC(=O)C(CS)NC(=O)C(CC5=CNC=N5)NC(=O)C6CCCN6C(=O)C(C)NC(=O)CNC(=O)C(CC(=O)O)NC(=O)C(C)NC(=O)C(CO)NC(=O)C(CC(=O)O)NC(=O)C(CC(C)C)NC(=O)CNC(=O)C(C)NC(=O)C(C)NC(=O)CNC(=O)C7CCCN7C(=O)C(CC(C)C)NC(=O)C(CS)NC(=O)C(CCSC)NC(=O)C(CC(=O)N)NC(=O)C(C)NC(=O)C(CCC(=O)O)NC(=O)C(CC(C)C)NC(=O)C(CC(=O)O)NC(=O)C(CC(C)C)NC(=O)C(CC8=CNC9=CC=CC=C98)NC(=O)C(CO)NC(=O)C(CC(C)C)NC(=O)C(CC(=O)O)NC(=O)C(CCCNC(=N)N)NC(=O)C1CCCN1C(=O)C(C(C)C)NC(=O)C(CS)NC(=O)C(CC1=CNC=N1)NC(=O)C1CCCN1C(=O)C(C)NC(=O)CNC(=O)C(CC(=O)O)NC(=O)C(C)NC(=O)C(CO)N

1. LRRK2 dephosphorylation increases its ubiquitination

Jing Zhao, R Jeremy Nichols, Tyler P Molitor, J William Langston Biochem J . 2015 Jul 1;469(1):107-20. doi: 10.1042/BJ20141305.

Activating mutations in the leucine rich repeat protein kinase 2 (LRRK2) gene are the most common cause of inherited Parkinson's disease (PD). LRRK2 is phosphorylated on a cluster of phosphosites including Ser(910), Ser(935), Ser(955) and Ser(973), which are dephosphorylated in several PD-related LRRK2 mutants (N1437H, R1441C/G, Y1699C and I2020T) linking the regulation of these sites to PD. These serine residues are also dephosphorylated after kinase inhibition and lose 14-3-3 binding, which serves as a pharmacodynamic marker for inhibited LRRK2. Loss of 14-3-3 binding is well established, but the consequences of dephosphorylation are only now being uncovered. In the present study, we found that potent and selective inhibition of LRRK2 kinase activity leads to dephosphorylation of Ser(935) then ubiquitination and degradation of a significant fraction of LRRK2. GNE1023 treatment decreased the phosphorylation and stability of LRRK2 in expression systems and endogenous LRRK2 in A549 cells and in mouse dosing studies. We next established that LRRK2 is ubiquitinated through at least Lys(48) and Lys(63) ubiquitin linkages in response to inhibition. To investigate the link between dephosphorylation induced by inhibitor treatment and LRRK2 ubiquitination, we studied LRRK2 in conditions where it is dephosphorylated such as expression of PD mutants [R1441G, Y1699C and I2020T] or by blocking 14-3-3 binding to LRRK2 via difopein expression, and found LRRK2 is hyper-ubiquitinated. Calyculin A treatment prevents inhibitor and PD mutant induced dephosphorylation and reverts LRRK2 to a lesser ubiquitinated species, thus directly implicating phosphatase activity in LRRK2 ubiquitination. This dynamic dephosphorylation-ubiquitination cycle could explain detrimental loss-of-function phenotypes found in peripheral tissues of LRRK2 kinase inactive mutants, LRRK2 KO (knockout) animals and following LRRK2 inhibitor administration.

2. Targeting 14-3-3 protein, difopein induces apoptosis of human glioma cells and suppresses tumor growth in mice

Weidong Cao, Jie Zhou, Lei Cao, Xiang Zhang, Zenghui Teng, Zhou Fei, Xiaoliang Yang Apoptosis . 2010 Feb;15(2):230-41. doi: 10.1007/s10495-009-0437-4.

14-3-3 protein has emerged as critical regulators of diverse cellular responses. Previous studies found that strong 14-3-3 protein expression was observed and associated with tumor genesis and progression in glioma. Here, we further elucidated the role of 14-3-3 protein in apoptosis of human glioma U251 and U87 cells by global inhibition of 14-3-3 functions with a general 14-3-3 antagonist, difopein. In vitro, morphological observation and DNA laddering assay showed that difopein-treated glioma cells displayed outstanding apoptosis characteristics, such as nuclear fragmentation, appearance of membrane-enclosed apoptotic bodies and DNA laddering fragment. Moreover, flow cytometric detection of phosphatidylserine externalization indicated that difopein-induced apoptosis occurred in a time-dependent manner. Interestingly, inhibiting 14-3-3 with small interfere RNA also induce apoptosis of human glioma U251 cells. Furthermore, RT-PCR and western blot assay further substantiated that difopein had strong effects to induce glioma cell apoptosis through down-regulating Bcl-2, up-regulating Bax and activating caspase-9 and caspase-3. In vivo, retroviral vector was constructed and retroviral-mediated transfer of difopein to glioma was implanted in nude mice. Difopein effectively hindered proliferation and triggered apoptosis of tumor cells implanted into nude mice. This work not only reveals a critical role of 14-3-3 in apoptosis suppression in glioma cells, but also identifies and validates 14-3-3 as a potential molecular target for anticancer therapeutic development.

3. Implication of 14-3-3ε and 14-3-3θ/τ in proteasome inhibition-induced apoptosis of glioma cells

Chao Li, Ying Yan, Yan-Yan Gao, Qiang Zhang, Hua-Qin Wang, Ying Xu, Zhi-Hong Zong Cancer Sci . 2013 Jan;104(1):55-61. doi: 10.1111/cas.12033.

Proteasome inhibitors represent a novel class of anticancer agents that are used in the treatment of hematologic malignancies and various solid tumors. However, mechanisms underlying their anticancer actions were not fully understood. It has been reported that strong 14-3-3 protein expression is observed and associated with tumor genesis and progression of astrocytoma. In addition, global inhibition of 14-3-3 functions with a general 14-3-3 antagonist difopein induces apoptosis of human astrocytoma cells, validating 14-3-3 as a potential molecular target for anticancer therapeutic management. In the current study, for the first time we demonstrated that proteasome inhibitors downregulated 14-3-3ε and 14-3-3θ/τ in U87 and SF295 glioma cells. Overexpression of 14-3-3ε and 14-3-3θ/τ significantly suppressed apoptosis of human glioma cells induced by proteasome inhibitors. We also demonstrated that MG132 activated ASK1 and siASK1 compromised the MG132-induced apoptosis of glioma cells. Furthermore, overexpression of 14-3-3ε and 14-3-3θ/τ markedly suppressed activation of ASK1. Collectively, the current study supported that proteasome inhibitors, at least in part, caused cytotoxicity of glioma cells via downregulation of 14-3-3ε and 14-3-3θ/τ and subsequent activation of ASK1.