Erythropoietin/CD131 heteroreceptor

Cibinetide (ARA290) promotes endothelial colony-forming cells' (ECFCs') motility, proliferation, and resistance against H2O2-induced apoptosis[1]. Cibinetide (ARA290) is an EPO-analog peptide that may have neurotrophic and depressive properties but has no hematological side effects[2].

---

ARA290 increases ECFC survival after oxidative stress in vitro by an anti-apoptotic effect [1]
ARA290 significantly reduced LDH release from H2O2-exposed ECFCs at 1 ng/mL (70.4% (53.3; 77.5), P < 0.05), 100 ng/mL (72.3% (52.1; 79.9), P < 0.01), and 1000 ng/mL (77.1% (53.8; 86.8), P < 0.05; Fig. 1A) compared with control cells. ARA290 treatment enhanced survival in a comparable manner than EPO 5 IU/mL treatment, used as a positive control (67.6% (59.6; 79.8) P < 0.01) compared with control.

---

---

ARA290 increases ECFC proliferation and migration in vitro [1]
ARA290 significantly increased proliferation of ECFCs in culture as shown by BrdU incorporation at concentration 100 ng/mL (136.4% (111.6; 170.0), P < 0.01) and 1000 ng/mL 128.4% (116.3; 151.4), P < 0.05; Fig. 2A). Proliferation increased after ARA290 treatment is comparable to EPO 5 UI/mL treatment used as a positive control (136.8% (127.2; 147.5), P < 0.01 vs. Ctrl). The increase proliferation induced by ARA290 1 ng/mL (125.1% (116.3; 123.9) was not significant (P = 0.1).

---

ARA290 Improves β Cell Secretory Function in GK Islets [2]
To assess whether ARA290 exerts a direct effect on β-cell secretory function, which may account for the improved glucose homeostasis in GK rat, we assessed effects of ARA290 on insulin secretion by performing GSIS experiments on islets isolated after 4 wks of ARA290 treatment in W and GK rats as well as batch incubations and islets perifusion experiments. In islets from ARA290-treated GK rats, insulin responses to 16.7 mmol/L glucose, in relation to basal (3.3 mmol/L) glucose, were significantly increased about two-fold compared with responses in islets from the placebo-treated rats, the fold increase in response being 3.8 ± 0.5 versus 2.0 ± 0.4, respectively (p < 0.05).
Islets isolated from untreated W and GK rats were exposed to ARA290 in the range of 1 to 10 ng/mL at both 3.3 and 16.7 mmol/L glucose. In W rat islets, ARA290 did not change insulin secretion during either basal or high glucose stimulation (Figure 2A). In GK rat islets, exposure to ARA290 at basal glucose conditions did not enhance insulin secretion, although in the presence of the high glucose concentration, 1 ng/mL ARA290 significantly improved insulin secretion 2.7-fold compared with 16.7 mmol/L glucose alone (Figure 2B). Higher concentrations of ARA290 further increased insulin secretion.
Islet perifusion experiments showed that ARA290 greatly augmented the first phase insulin response (control 0.076 ± 0.017 versus ARA290 0.303 ± 0.184 μU/islet/min at 34 min, p < 0.01) (Figure 2C). By contrast, ARA290 did not exert a significant effect on the second phase insulin secretion. However, addition of KCl and ARA290 increased insulin secretion further (Figure 2C), suggesting that ARA290 affects not only the glucose stimulatory pathway but also the amplification pathway.

---

ARA290 Improves Islet Glucose Metabolism [2]
To address whether improved first phase insulin secretion could result from a beneficial effect of A290 on glucose metabolism, we assessed effects of ARA290 on islet glucose oxidation and ATP production. Under hyperglycemic conditions, ARA290 significantly increased glucose oxidation (Figure 3A) in GK islets. Treatment by ARA290 for 1 h also improved ATP production in GK islets (Figure 3B). These results suggest that ARA290 has a direct effect on glucose metabolism by increasing Krebs cycle activity and ATP production by the mitochondria.

---

ARA290 Augments the Insulin Secretion Pathway [2]
To investigate whether ARA290 has effects on the islet KATP channel in the insulin secretion pathway, we used the KATP channel opener diazoxide. At 3.3 mmol/L glucose, diazoxide did not suppress basal insulin secretion, and the stimulation of insulin secretion by addition of diazoxide and KCl was not significantly modulated by ARA290 (Figure 4A). However, in the presence of 16.7 mmol/L glucose, coincubation of GK rat islets with 10 ng/mL ARA290 and 0.25 mmol/L diazoxide neutralized the stimulatory effect of ARA290 on GSIS (Figure 4B). This suggests that the stimulatory effect on GSIS by ARA290 is mediated through KATP channels. However, at high glucose concentrations when the KATP channels were kept open by diazoxide and were depolarized by 30 mmol/L KCl, ARA290 potentiated the GSIS of GK islets (Figure 4B). These results show that ARA290 has an additional effect in the insulin secretion pathway that is distal of the KATP channels. Indeed, when GK islets were coincubated with 10 ng/mL ARA290 and 10 μmol/L of the protein kinase A (PKA) inhibitor H89, the effect of ARA290 on GSIS was abolished (Figure 4C). Altogether these results suggest that ARA290 exerts its effect on GSIS both on the KATP channels and on the insulin amplification pathway through PKA. Finally, we examined whether the effect of ARA290 was dependent on Ca2+ signaling. The effect of ARA290 on GSIS was blocked by the addition of the L-type Ca channel blocker nimodipine (5 μmol/L) (Figure 4D). This suggests that improvement of insulin secretion by ARA290 involves an entry of Ca2+ through the L-type Ca2+ channels.

A single Cibinetide (ARA290) injection improves the hindlimb blood flow and capillary density after 28 days of ECFC transplantation in mice with CLI. It also improves the homing of radiolabeled transplanted cells to the ischemic leg 4 hours after transplantation[1]. Rats' body weight is unaffected by cipinetide (ARA290; 30 μg/kg, sc), which inhibits the increasing deterioration of glucose regulation. In GK rats, cipinetide dramatically lowers glucose AUCs in IPGTT[2]. Rats treated with low-dosage Cibinetide (35 μg/kg, ip) have just a small reduction in the severity of EAE. In a dose-dependent manner, the group receiving curenetide treatment (70 μg/kg, ip) considerably postpones the onset, reduces the neurologic severity, and shortens the duration of EAE[3].

---

Enhanced angiogenic response of ARA290 and ECFC co-administrated in vivo [1]
Recovery of blood flow in the ischemic hindlimb was significantly increased in ARA29010 μg/kg (day 28: 61.6 ± 4.2%, P < 0.05), ECFC (day 28: 65.4 ± 2.3%, P < 0.05), and ECFC + ARA29010 μg/kg groups (day 28: 79.2 ± 4.6%, P <0.001) as compared with control group (day 28: 53.0 ± 4.1%) (Fig. 3A). A two-way ANOVA analysis followed by a Bonferroni test on day 7 post-injury revealed that the combination therapy showed an earlier improvement in hindlimb blood perfusion (ECFC + ARA29010 μg/kg: 74.5 ± 3.6%) as compared with ARA29010 μg/kg group (ARA29010 μg/kg: 54.0 ± 2.29 %, P < 0.001) or ECFCs group (ECFC: 46.9 ± 3.1%, P <0.001) alone or control group (50.6 ± 1.36%, P < 0.001).
On day 28 post-injury, ECFC + ARA29010 μg/kg group showed higher ischemic/non-ischemic capillary density ratio (1.255 (1.16; 1.39)) in gastrocnemius muscle compared with control group (0.66 (0.57; 0.88); P < 0.001), ARA29010 μg/kg group (0.87 (0.73; 0.97); P < 0.01), or ECFC group (1.02 (0.8; 1.15); P < 0.01) (Fig. 3B). Moreover, ECFC alone produced a higher ratio than control group (P < 0.01); and ARA29010 μg/kg alone tends to increase this ratio (P = 0.13).
On day 7 post injury, the clinical necrosis score was significantly decreased in ECFC (0 (0; 1), P < 0.05), ARA29010 μg/kg (0 (0; 1), P < 0.05) and ECFC + ARA29010 μg/kg (0 (0; 1), P < 0.05) groups as compared with control group (1 (1; 6) (Fig. 3C). Although ANOVA is non-significant, mobility impairment at day 7 tends to be lower in ECFC (0 (0; 2)), ARA29010 μg/kg (0 (0; 3)), and ECFC + ARA29010 μg/kg (0 (0; 2)) groups than in control group (2 (0; 4)).

---

ARA290-induced CD31 up-regulation in ECFCs that participate in homing enhancement of ECFC to ischemic site in vivo [1]
Focusing on the hindlimbs region of interest, ARA290 co-administrated with ECFC increased the transplanted cells homing to the ischemic limb, as shown by ischemic/non-ischemic count ratio (163.2% (139.3; 185.0), P < 0.001) compared with ECFC alone (96.2% (50.5; 102.0)).
Western blot experiments revealed a CD31-identified 120 kDa to 130 kDa protein in whole cell lysate of ECFCs. Expression levels of CD31 plotted as percentage of control ECFCs were significantly increased after a 4-h incubation period with ARA290 at 1 ng/mL: 146.0% (121.6; 205.2), P < 0.05; and 100 ng/mL: 147.4% (116.6; 211.9), P < 0.05; and a positive trend with 1000 ng/mL: 129.0% (110.2; 174.3). P = 0.07). EPO treatment produced a non-significant increase (135.9% (100.2; 174.0), NS) (Fig. 4A).
When ECFC were incubated with an anti-CD31 blocking antibody before transplantation, we observed a significant decrease in the ischemic/non-ischemic 99mTc-ECFC ratio in ECFC + ARA29010 μg/kg (93.68% (28.0; 115.0), P < 0.001) compared with non-incubated cells, and a trend to decrease the ischemic/non-ischemic 99mTc-ECFC ratio in ECFC group (34.0% (16.3; 76.6), P = 0.06) (Fig. 4B).

---

Effects of ARA290 on glucose homeostasis were studied in type 2 diabetic Goto-Kakizaki (GK) rats. In GK rats receiving ARA290 daily for up to 4 wks, plasma glucose concentrations were lower after 3 and 4 wks, and hemoglobin A1c (Hb A1c) was reduced by ~20% without changes in whole body and hepatic insulin sensitivity. Glucose-stimulated insulin secretion was increased in islets from ARA290-treated rats. Additionally, in response to glucose, carbachol and KCl, islet cytoplasmic free Ca2+ concentrations, [Ca2+]i, were higher and the frequency of [Ca2+]i oscillations enhanced compared with placebo. ARA290 also improved stimulus-secretion coupling for glucose in GK rat islets, as shown by an improved glucose oxidation rate, ATP production and acutely enhanced glucose-stimulated insulin secretion. ARA290 also exerted an effect distal to the ATP-sensitive potassium (KATP) channel on the insulin exocytotic pathway, since the insulin response was improved following islet depolarization by KCl when KATP channels were kept open by diazoxide. Finally, inhibition of protein kinase A completely abolished effects of ARA290 on insulin secretion. In conclusion, ARA290 improved glucose tolerance without affecting hematocrit in diabetic GK rats. This effect appears to be due to improved γ-cell glucose metabolism and [Ca2+]i handling, and thereby enhanced glucose-induced insulin release. [2]

---

ARA290 is a nonerythropoietic analog of erythropoietin (EPO) containing 11 amino acids which provides the anti-inflammatory and neuroprotective effects of EPO without stimulating hematopoiesis. Here we studied the therapeutic effects of ARA290 in experimental autoimmune encephalomyelitis (EAE) Lewis rats. Therapeutic (from Day 7 to Day 18 or from Day 9 to Day 19) administration of ARA290 (35, 70 μg/kg, intra-peritoneal) to EAE rats once daily significantly reduced the severity and shortened the duration of clinical score, reduced the accumulation of inflammatory cells in EAE spinal cords and suppressed mRNA levels of interleukin-1β (IL-1β), IL-17, tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), inducible nitric oxide synthase (iNOS), matrix metalloproteinase 9 (MMP9) and transcription factor T-bet in spinal cords of EAE rats. Furthermore, ARA290 treatment reduced the helper T cell number in lymph nodes and circulation in EAE. In vitro study showed that ARA290 dose-dependently inhibited antigen specific- and antigen non-specific-lymphocyte proliferation as well. In addition, ARA290 altered the cytokine milieu to favor the polarization of Th2 and regulatory T (Treg) cells but suppressed the polarization of Th1 and Th17 cells in EAE lymph nodes. In summary, our study here showed that ARA290 could alter T cell function to suppress inflammation to ameliorate EAE, suggesting that ARA290 may be a new therapeutic candidate for multiple sclerosis[3].

Batch-Incubation Experiments [2]
After the overnight culture, the islets were preincubated for 30 min at 37°C in Krebs-Ringer bicarbonate buffer solution (KRB), supplemented with 2 mg/mL bovine albumin, 10 mmol/L HEPES and 3.3 mmol/L glucose, pH 7.4. After preincubation, batches of three islets were incubated for 60 min in a shaking water bath at 37°C in 300 μL of KRB with either 3.3 or 16.7 mmol/L glucose. In both conditions, islets were treated with or without addition of 1, 5 or 10 ng/mL ARA290. When effects of ARA290 on the insulin secretion pathway was analyzed, KRB with either 3.3 mmol/L or 16.7 mmol/L glucose was supplemented with 0.25 mmol/L diazoxide ± 30 mmol/L KCl to analyze proximal or distal effect of ARA290 on the ATP-sensitive potassium (KATP) channels, or with 10 μmol/L PKA inhibitor H89, or 5 μmol/L nimodipine to block the L-type of Ca2+ channels. After incubations, aliquots of the media were taken for radioimmunoassay of insulin (11).

---

Islets Perifusion [2]
After overnight culture in RPMI, GK rat islets were preincubated for 30 min in KRB buffer supplemented with 3.3 mmol/L glucose. Batches of 50 islets were layered between two layers of bio-gel (Bio-Rad) in a perifusion chamber and perifused at a flow rate of 200 μL/min, at 37°C, with KRB and 3.3 mmol/L glucose for 20 min prior to the start of collecting samples. To determine the dynamic response of insulin secretion, the islets were first perifused for 20 min with 3.3 mmol/L glucose followed by 10 min with 3.3 mmol/L glucose plus 10 ng/mL ARA290, then for 30 min with 16.7 mmol/L glucose in absence or presence of 10 ng/mL ARA290, then for 15 min KCl 30 mmol/L in absence or presence of 10 ng/mL ARA290, followed by 30 min 3.3 mmol/L glucose only. Samples were collected every 2 min and stored at −20°C until insulin was analyzed by radioimmunoassay.

---

Glucose Oxidation [2]
After overnight culture in RPMI, GK rat islets were preincubated for 30 min in KRB buffer supplemented with 3.3 mmol/L glucose. Ten islets were placed in glass incubation vials with either 3.3 mmol/L or 16.7 mmol/L glucose in 100 μL KRB (pH 7.4), with or without 10 ng/mL ARA290 and also containing 1 μCi D-[U-14C]glucose). Incubation vials were then placed in 20 mL scintillation bottles containing 1.5 mL water and sealed with a rubber-membrane equipped cap under 95% O2, 5% CO2 and incubated for 2 h in 37°C in a water bath. The incubation was terminated by injecting through the rubber membrane 100 μL of 0.05 mmol/L antimycin in 70% ethanol, followed by 250 μL of hyamine in the scintillation bottles and 100 μL of 0.4 mmol/L sodium phosphate buffer, pH 6.0, in the incubation vials. The 14CO2 was allowed to absorb overnight into the hyamine. Incubation vials were discarded and 5 mL of scintillation liquid were added in the scintillation bottles. Radioactivity in 14CO2 was measured with a Liquid Scintillator Analyzer, and results were expressed as pmoles glucose oxidized/islet per 2 h.

---

ATP Determination [2]
After preincubation as above, batches of 20 islets were incubated for 1 h in 300 μL KRB with either 3.3 mmol/L or 16.7 mmol/L glucose, and with or without 10 ng/mL ARA290, at 37°C in a shaking water bath. ATP levels were measured using the ATP Bioluminescence Assay Kit HS II according to the manufacturer’s instructions. Bioluminescence was measured after 1-s delay and with 1-s integration time, using a GloMax 96 Microplate Luminometer. Protein concentration in the lysis suspension was determined by the Bradford protein assay (Bio-Rad), and results were expressed as pmol ATP/mg protein.

In vitro experiments [1]
For all experiments, ARA290 was tested at different doses: 1 ng/mL, 100 ng/mL, and 1000 ng/mL. EPO 5 UI/mL has been used as a positive control.

---

Survival assays [1]
ECFC survival after oxidative stress was measured by the LDH release assay. Released LDH due to cell lysis was measured using cytotoxicity detection kit according to the manufacturer recommendations. Briefly, ECFCs were plated in 96-well plates and treatment was added to well's control medium for priming during 24 h. Primed ECFCs were subsequently washed and exposed to 0.5% FBS/EBM-2 with H2O2 1100 μM for 18 h. Then, supernatants were incubated with kit reagents and incubated at 20°C for 30 min. Absorbance at 492 nm was measured using an ELISA plate reader . The average absorbance values were normalized by subtracting mean culture medium absorbance values. Results were expressed as percentage of H2O2-treated control cells, n= 3 to 4 per group. [1]
The effect of ARA290 on H2O2-induced apoptosis of ECFCs was determined using the 7AAD-Annexin V-FITC kit (Beckman Coulter, Villepinte, France). Samples were read in a FC500 flow cytometer. Cells that showed low staining for both Annexin V and 7-AAD were considered prosurvival cells. Cells with high Annexin V staining regardless of 7-AAD status are considered apoptotic cells. Results were expressed as percentage of H2O2-treated control cells, n = 4 per group.

---

BrdU incorporation [1]
ECFC proliferation was measured by quantitating bromodeoxyuridine (BrdU) incorporated into the newly synthesized DNA of replicating ECFC using the Cell Proliferation ELISA BrdU assay, according to the manufacturer's instructions. Briefly, 1 × 10~4 cells/well of ECFC were seeded and incubated in 96-well plastic plates in EGM-2 and cultured for 48 h. Then, BrdU labeling reagent was added in culture medium 24 h before assay in the presence or not of ARA290 (1–1,000 ng/mL). Then cells were denaturated and cell lysates were incubated for 90 min with 1:100 diluted mouse anti-BrdU mAb conjugated to peroxidase, washed and substrate solution was added for 20 min. Finally, the absorbance was measured at 405 nm with a reference wavelength at 530 nm using an ELISA plate reader. Results were expressed as percentage of control cells, n = 4 per group.

Diabetic Goto-Kakizaki (GK) rats, originating from Wistar rats, are bred in our department. Normal Wistar (W) rats are used as nondiabetic controls. All animals are about six weeks old and with body weights 100 to 150 g when treatment is initiated. They are kept at 22°C on a reversed 12-h light-dark cycle with free access to food, except when fasted overnight as noted below. Rats are treated over 4 wks with Cibinetide (ARA290) by a once daily subcutaneous (s.c.) injection at a dose of 30 μg/kg body weight or PBS. Blood samples for determination of glucose are taken after a small tail incision and analyzed every week before morning s.c. injection of either ARA290 or placebo. During the experimental period, body weights are measured weekly.

---

Mouse model of hindlimb ischemia [1]
For surgical procedures, animals were anesthetized with 1.5 vol% isoflurane in a gas mixture of 30% oxygen and 70% nitrogen and placed on a homeothermic blanket. Local analgesia bupivacaine (5 mg/kg) was injected along the incision. Unilateral hindlimb ischemia was performed after femoral artery excision as previously described. After 24 h mice were randomly allocated into treatment groups; intravenous injections of PBS (Ctrl) (n = 6), 105 ECFCs per mouse (ECFC) (n = 7), 10 μg/kg Cibinetide (ARA290) (ARA290,10 μg/kg) (n = 7), or 105 ECFCs per mouse + 10 μg/kg (ECFC + ARA290,10 μg/kg) (n = 7). Cibinetide (ARA290) dosing was determined as a single injection of 10 μg/kg because it demonstrated protective effects in a previous report and a single dose of EPO was sufficient to enhance ECFCs transplantation-induced angiogenesis in a model of transient focal cerebral ischemia. Laser Doppler perfusion imaging was used to assess revascularization from day 1 to day 28 after surgery. Perfusion results are expressed as a ratio of ischemic to non-ischemic limb blood flow. Hindlimb ischemic damage was quantified on day 7, as previously described. Briefly, the motility impairment score was calculated for each mouse as follows: 0, unrestricted active movement; 2, full use of the foot but no spreading toes; 3, restricted active foot, no plantar flexion; 4, dragging foot. The necrosis score was calculated for each mouse as follows: 0, no necrosis; 1, foot oedema; 2, necrosis of one toe; 3, necrosis of two or more toes; 4, foot necrosis; 5, leg necrosis; and 6, autoamputation of the entire leg. Scores were ascribed in a randomized and blinded fashion by trained experimenter.

---

The nonhematopoietic erythropoietin analogue Cibinetide (ARA290) consists of 11 amino acids (MW 1258 daltons), and was supplied by Araim Pharmaceuticals. It was dissolved in phosphate buffered saline (PBS) at a concentration of 2 mg/mL and kept at 4°C for up to 4 wks.
Diabetic Goto-Kakizaki (GK) rats, originating from Wistar rats, were bred in our department. Normal Wistar (W) rats were purchased from a commercial breeder and used as nondiabetic controls. All animals were about six weeks old and with body weights 100 to 150 g when treatment was initiated. They were kept at 22°C on a reversed 12-h light–dark cycle with free access to food, except when fasted overnight as noted below. All in vivo experiments were performed in a blinded manner. Rats were treated over 4 wks with Cibinetide (ARA290) by a once daily subcutaneous (s.c.) injection at a dose of 30 μg/kg bodyweight or PBS. Blood samples for determination of glucose were taken after a small tail incision and analyzed every week before morning s.c. injection of either Cibinetide (ARA290) or placebo. During the experimental period, body weights were measured weekly.[2]

[1]. ARA290, a Specific Agonist of Erythropoietin/CD131 Heteroreceptor, Improves Circulating Endothelial Progenitors' Angiogenic Potential and Homing Ability. Shock. 2016 Oct;46(4):390-7.

[2]. ARA290 Improves Insulin Release and Glucose Tolerance in Type 2 Diabetic Goto-Kakizaki Rats. Mol Med. 2015; 21(1): 969–978.

[3]. Therapeutic effects of nonerythropoietic erythropoietin analog ARA290 in experimental autoimmune encephalomyelitis rat. J Neuroimmunol. 2014 Mar 15;268(1-2):64-70.

Cibinetide has been used in trials studying the basic science of Depression.

---

Drug Indication
Treatment of sarcoidosis

---

Background: Alternate erythropoietin (EPO)-mediated signaling via the EPOR/CD131 heteromeric receptor exerts the tissue-protective actions of EPO in a wide spectrum of injuries, especially ischemic diseases. Circulating endothelial progenitor cells contribute to endothelial repair and post-natal angiogenesis after chronic ischemic injury. This work aims to investigate the effects of ARA290, a specific agonist of EPOR/CD131 complex, on a subpopulation of endothelial progenitor cells named endothelial colony-forming cells (ECFCs) and to characterize its contribution to ECFCs-induced angiogenesis after peripheral ischemia. Methods: ARA290 effects on ECFCs properties were studied using cell cultures in vitro. We injected ARA290 to mice undergoing chronic hindlimb ischemia (CLI) in combination with ECFC transplantation. The homing of transplanted ECFC to ischemic tissue in vivo was assessed by SPECT/CT imaging. Results: In vitro, ARA290 enhanced the proliferation, migration, and resistance to H2O2-induced apoptosis of ECFCs. After ECFC transplantation to mice with CLI, a single ARA290 injection enhanced the ischemic/non-ischemic ratio of hindlimb blood flow and capillary density after 28 days and the homing of radiolabeled transplanted cells to the ischemic leg 4 h after transplantation. Prior neutralization of platelet-endothelial cell adhesion molecule-1 (CD31) expressed by the transplanted cells inhibited ARA290-induced improvement of homing. Discussion: ARA290 induces specific improvement of the biological activity of ECFCs. ARA290 administration in combination with ECFCs has a synergistic effect on post-ischemic angiogenesis in vivo. This potentiation appears to rely, at least in part, on a CD31-dependent increase in homing of the transplanted cells to the ischemic tissue. [1]

C51H84N16O21

1257.32

1256.599

C, 48.72; H, 6.73; N, 17.82; O, 26.72

1208243-50-8

1208243-50-8;Cibinetide acetate;

91810664

{Glp}-Glu-Gln-Leu-Glu-Arg-Ala-Leu-Asn-Ser-Ser; H-Pyr-Glu-Gln-Leu-Glu-Arg-Ala-Leu-Asn-Ser-Ser-OH; L-pyroglutamyl-L-alpha-glutamyl-L-glutaminyl-L-leucyl-L-alpha-glutamyl-L-arginyl-L-alanyl-L-leucyl-L-asparagyl-L-seryl-L-serine

{Glp}-EQLERALNSS; XEQLERALNSS

Typically exists as solid at room temperature

1.6±0.1 g/cm3

1.661

-6.59

20

22

42

88

2560

11

O=C([C@]([H])(C([H])([H])C([H])(C([H])([H])[H])C([H])([H])[H])N([H])C([C@]([H])(C([H])([H])C([H])([H])C(N([H])[H])=O)N([H])C([C@]([H])(C([H])([H])C([H])([H])C(=O)O[H])N([H])C([C@]1([H])C([H])([H])C([H])([H])C(N1[H])=O)=O)=O)=O)N([H])[C@]([H])(C(N([H])[C@]([H])(C(N([H])[C@@]([H])(C([H])([H])[H])C(N([H])[C@]([H])(C(N([H])[C@]([H])(C(N([H])[C@]([H])(C(N([H])[C@]([H])(C(=O)O[H])C([H])([H])O[H])=O)C([H])([H])O[H])=O)C([H])([H])C(N([H])[H])=O)=O)C([H])([H])C([H])(C([H])([H])[H])C([H])([H])[H])=O)=O)C([H])([H])C([H])([H])C([H])([H])/N=C(\N([H])[H])/N([H])[H])=O)C([H])([H])C([H])([H])C(=O)O[H]

WZTIQQBMSJTRBR-WYKNNRPVSA-N

InChI=1S/C51H84N16O21/c1-22(2)17-30(47(84)65-32(19-36(53)71)48(85)66-33(20-68)49(86)67-34(21-69)50(87)88)63-40(77)24(5)57-41(78)25(7-6-16-56-51(54)55)59-43(80)29(11-15-39(75)76)62-46(83)31(18-23(3)4)64-45(82)27(8-12-35(52)70)60-44(81)28(10-14-38(73)74)61-42(79)26-9-13-37(72)58-26/h22-34,68-69H,6-21H2,1-5H3,(H2,52,70)(H2,53,71)(H,57,78)(H,58,72)(H,59,80)(H,60,81)(H,61,79)(H,62,83)(H,63,77)(H,64,82)(H,65,84)(H,66,85)(H,67,86)(H,73,74)(H,75,76)(H,87,88)(H4,54,55,56)/t24-,25-,26-,27-,28-,29-,30-,31-,32-,33-,34-/m0/s1

(4S)-5-[[(2S)-5-amino-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-4-amino-1-[[(2S)-1-[[(1S)-1-carboxy-2-hydroxyethyl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-1,4-dioxobutan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-1-oxopropan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-4-carboxy-1-oxobutan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-1,5-dioxopentan-2-yl]amino]-5-oxo-4-[[(2S)-5-oxopyrrolidine-2-carbonyl]amino]pentanoic acid

ARA-290; ARA 290; Cibinetide; 1208243-50-8; ARA290; PHBSP; ARA-290; PH-BSP; ARA 290; Cibinetide [USAN]

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 : ~66.67 mg/mL (~53.03 mM)
0.1 M NaOH :≥ 50 mg/mL (~39.77 mM)

Solubility in Formulation 1: 2.5 mg/mL (1.99 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution.

---

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