guanylate cyclase-C ( EC50 = 190 nM )
Plecanatide (1 nM-10 μM) stimulates the synthesis of cyclic guanosine monophosphate (cGMP) in a dose-dependent manner with an EC50 of 190 nM by activating the GC-C receptor[1].

Plecanatide (1 nM-10 μM) stimulates the synthesis of cyclic guanosine monophosphate (cGMP) in a dose-dependent manner with an EC50 of 190 nM by activating the GC-C receptor[1].

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Plecanatide stimulated cyclic GMP (cGMP) synthesis in human colonic carcinoma T84 cells in a dose-dependent manner, with an EC₅₀ value of 1.9 × 10⁻⁷ mol/L.[1]

Plecanatide (0.5 and 2.5 mg/kg, p.o.) improves colitis that is chemically and spontaneously induced in BALB/c mice after 7 days of treatment, and in TCRα-/- mice, it takes 14 days[1].
Plecanatide (0.005–5 mg/kg, once daily for seven days) also exhibits anti-inflammatory properties in BDF-1 mice that have colitis induced by trinitrobenzene sulfonic (TNBS) and dextran sulfate sodium (DSS)[1].

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Oral administration of plecanatide (0.05–0.5 mg/kg/day) ameliorated colitis in both dextran sulfate sodium (DSS)-induced and trinitrobenzene sulfonic acid (TNBS)-induced acute colitis models in mice, as evidenced by reduced colitis severity scores and disease activity index (DAI).[1]
Plecanatide (0.5 and 2.5 mg/kg/day) also reduced colitis severity in TCRα⁻/⁻ mice, a model of spontaneous chronic colitis.[1]
The anti-inflammatory effect of plecanatide was comparable to that of sulfasalazine (80 mg/kg) and 5-aminosalicylic acid (100 mg/kg) in DSS-induced colitis.[1]

T84 cells were pre-incubated with a phosphodiesterase inhibitor, followed by incubation with plecanatide. Intracellular cGMP levels were measured using an ELISA kit after reaction termination and neutralization.[1]

Female BALB/c mice (2-4 month old) are induced colitis by TNBS
0, 0.5 and 2.5 mg/kg
P.o. for 7 days

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In DSS-induced colitis in BDF1 mice, plecanatide (0.005–5 mg/kg) was administered daily by oral gavage in phosphate-buffered saline, starting one day before DSS administration and continuing for 7 days.[1]
In TNBS-induced colitis in BALB/c mice, plecanatide (0.5 and 2.5 mg/kg) was administered orally for 7 days starting on the day of TNBS challenge.[1]
In TCRα⁻/⁻ mice, plecanatide (0.5 and 2.5 mg/kg) was administered orally for 14 days.[1]
Colitis severity was assessed by histopathological scoring of H&E-stained colon sections and by calculating a disease activity index based on body weight, stool consistency, and fecal blood.[1]

Absorption, Distribution and Excretion
Following oral administration, pulcanatide is minimally absorbed, and systemic bioavailability is negligible. After oral administration of 3 mg, plasma concentrations of pulcanatide and its active metabolites are below the limit of quantitation. Therefore, standard pharmacokinetic parameters such as AUC, maximum concentration (Cmax), and half-life (t½) cannot be calculated. Human excretion studies have not been conducted. Pulcanatide and its active metabolites are undetectable in plasma after administration of the recommended clinical dose. After oral administration of 3 mg, plasma concentrations of pulcanatide and its active metabolites are below the limit of quantitation. Therefore, volume of distribution cannot be calculated. Human excretion studies have not been conducted. Given that the concentration of pulcanatide cannot be measured after clinically relevant oral doses, minimal tissue distribution of pulcanatide is expected. Orally administered pulcanatide is primarily distributed in the gastrointestinal tract, where it acts as a guanylate cyclase C (GC-C) agonist; systemic exposure is negligible. Prucanadide binds very little or almost no to human serum albumin or human α-1 acid glycoprotein. In a crossover study, 24 healthy subjects received a single 9 mg Trulance dose (3 times the recommended dose) under three different conditions: fasting; after a low-fat, low-calorie meal (LF-LC; approximately 350 calories: 17% fat, 66% carbohydrates, 17% protein); and after a high-fat, high-calorie meal (HF-HC; approximately 1000 calories: 60% fat, 25% carbohydrates, 15% protein). Prucanadide was detectable in one subject (fasting) at 0.5 hours and 1 hour after administration. At all other time points and in all other subjects, rucanadide concentrations were below the limit of quantitation. No active metabolites were detected in any subjects. Following oral administration of rucanadide, absorption is minimal, and systemic bioavailability is negligible. Following oral administration of 3 mg Trulance, plasma concentrations of rucanadide and its active metabolites were below the limit of quantitation in most analyzed plasma samples. Therefore, standard pharmacokinetic parameters such as AUC, maximum concentration (Cmax), and half-life cannot be calculated.

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Metabolism/Metabolites
Pucanatide is metabolized in the gastrointestinal tract to its active metabolite, which is achieved by the loss of a terminal leucine residue. Pucanatide and its metabolites are both degraded into smaller peptides and naturally occurring amino acids by proteases in the intestinal lumen.
Pucanatide is metabolized in the gastrointestinal tract to its active metabolite, which is achieved by the loss of a terminal leucine residue. Pucanatide and its metabolites are both degraded into smaller peptides and naturally occurring amino acids by proteases in the intestinal lumen.

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Biological Half-Life
Since systemic absorption is negligible, the half-life (t½) cannot be calculated.

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PucanatideIs hardly absorbed into the systemic circulation and mainly exerts its effects locally in the intestinal lumen. [1]

Toxicity Summary
Identification and Uses: Pucanatide is a white to off-white powder. Indications: For the treatment of chronic idiopathic constipation and constipation-predominant irritable bowel syndrome. Human Studies: Pucanatide is contraindicated in patients under 6 years of age and should be avoided in patients under 18 years of age. Animal Studies: In young mice, a single oral dose of pulcanatide resulted in dehydration and death. A 2-year carcinogenicity study was conducted in mice and rats to assess the carcinogenicity of pulcanatide. No tumorigenicity was observed in mice at oral doses up to 90 mg/kg/day, or in rats at oral doses up to 100 mg/kg/day. During organogenesis, pulcanatide was administered to pregnant mice and rabbits. No evidence of adverse effects on embryo-fetal development was found in mice at oral doses up to 800 mg/kg/day and in rabbits up to 250 mg/kg/day. In mice during organogenesis and lactation, oral doses up to 600 mg/kg/day showed no effects on offspring developmental abnormalities, growth, learning, memory, or fertility. In male or female mice, oral doses up to 600 mg/kg/day had no effect on fertility or reproductive function. In vitro bacterial reverse mutation (Ames) assays, in vitro mouse lymphoma mutation assays, and in vivo mouse bone marrow micronucleus assays showed no genotoxicity of purcanatide.
Hepatotoxicity
In premarketing clinical trials, the incidence of elevated serum enzymes during purcanatide treatment was low, with 1% to 2% of subjects experiencing some degree of serum ALT or AST elevation. Only 0.2% of patients had values exceeding 5 times the upper limit of normal. No clinically significant liver injury with jaundice associated with purcanatide treatment was found in these studies. Since the approval of purcanatide, there have been no published reports of elevated serum transaminases or clinically significant liver injury attributable to purcanatide. Therefore, liver injury caused by pucanatide, even if it occurs, is certainly very rare. Probability Score: E (Unlikely to be the cause of clinically significant liver injury). Pregnancy and Lactation Effects ◉ Overview of Use During Lactation: Pucanatide is not absorbed through the gastrointestinal tract. After a breastfeeding woman takes the recommended dose, the drug and its active metabolites are undetectable in breast milk. Pucanatide is not expected to have any adverse effects on breastfed infants. No special precautions are required. ◉ Effects on Breastfed Infants: As of the revision date, no relevant published information was found. ◉ Effects on Lactation and Breast Milk: As of the revision date, no relevant published information was found. Protein Binding Pucanatide binds very little or no to human serum albumin or human α-1-acid glycoprotein.

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In two phase III clinical trials for chronic idiopathic constipation (CIC), the most common treatment-emergent adverse event (TEAE) during treatment with purcanatide was diarrhea. [2]
In the first study (NCT01982240), the incidence of diarrhea was 5.9% and 5.7% in the 3 mg and 6 mg groups, respectively, compared to 1.3% in the placebo group. The rates of discontinuation due to diarrhea were 2.7% in the 3 mg group, 2.6% in the 6 mg group, and 0.4% in the placebo group.[2]
In the second study (NCT02122471), the incidence of diarrhea was 3.2% and 4.5% in the 3 mg and 6 mg groups, respectively, compared to 1.3% in the placebo group. The rate of discontinuation due to diarrhea was 1.1% in each purcanatide group and 0.4% in the placebo group.[2]
Most treatment-emergent adverse events (TEAEs) were mild to moderate. The incidence of serious TEAEs was ≤3.7% in all treatment groups. No deaths were reported. [2]
In the NCT02122471 study, a patient taking a 6 mg dose experienced an elevation in liver function parameters (ALT/AST), a serious adverse event (SAE) that was considered potentially related to the study drug. However, the patient was also taking indomethacin, and the elevation persisted after discontinuation of the drug. [2]

[1]. Plecanatide and dolcanatide, novel guanylate cyclase-C agonists, ameliorate gastrointestinal inflammation in experimental models of murine colitis. World J Gastrointest Pharmacol Ther. 2015 Nov 6;6(4):213-22.

[2]. Plecanatide: a new guanylate cyclase agonist for the treatment of chronic idiopathic constipation. Therap Adv Gastroenterol. 2018 Jun 8;11:1756284818777945.

[3]. Plecanatide, an oral guanylate cyclase C agonist acting locally in the gastrointestinal tract, is safe. Dig Dis Sci. 2013 Sep;58(9):2580-6.

Therapeutic Uses
Guanylate cyclase C agonist; Gastrointestinal medications. ClinicalTrials.gov is a registry and results database that lists human clinical studies funded by public and private institutions worldwide. The website is maintained by the National Library of Medicine (NLM) and the National Institutes of Health (NIH). Each record on ClinicalTrials.gov includes a summary of the study protocol, including: the disease or condition; the intervention (e.g., the medical product, behavior, or procedure under investigation); the title, description, and design of the study; participation requirements (eligibility criteria); the location of the study; contact information for the study location; and links to relevant information from other health websites, such as the NLM's MedlinePlus (for patient health information) and PubMed (for citations and abstracts of academic articles in the medical field). Prucanatide is listed in the database. Trulance is indicated for the treatment of: Chronic idiopathic constipation (CIC); Constipation-predominant irritable bowel syndrome (IBS-C). /Included in US Product Labelling/
/Exploratory Treatment/ /This study aims to/evaluate the ameliorative effect of oral purcanatide or docanatide (a guanosine analogue) on a mouse model of colitis. The cyclic guanosine monophosphate (cGMP) stimulatory potency of purcanatide and docanatide was determined using a human colon cancer T84 cell-based assay. In animal experiments, all test drugs were prepared in phosphate-buffered saline. Sulfasalazine or 5-aminosalicylic acid (5-ASA) served as positive controls. This study investigated the ameliorative effect of oral test drugs on acute colitis induced by drinking water supplementation with sodium dextran sulfate (DSS) or rectal instillation with trinitrobenzenesulfonic acid (TNBS) in BALB/c and/or BDF1 mice. Furthermore, the effect of oral purcanatide on spontaneous colitis in T-cell receptor α gene knockout (TCRa(-/-)) mice was also investigated. Colitis remission was assessed by monitoring colitis severity, disease activity index, and histopathology. Myeloperoxidase activity was measured using frozen colon tissue. Pucanatide and docanatide are structure-related guanosine analogs, guanosine being an endogenous ligand for guanylate cyclase C (GC-C). As expected of GC-C agonists, both pulcanatide and docanatide exhibited potent cGMP-stimulating activity in T84 cells. Once-daily gavage administration of these two analogs (0.05–0.5 mg/kg) improved colitis symptoms in DSS and TNBS-induced acute colitis models, as assessed by weight loss, reduced colitis severity (P < 0.05), and decreased disease activity index (P < 0.05). The ameliorative effects of both candidates on colitis were comparable to those of oral sulfasalazine or 5-aminosalicylic acid (5-ASA). Pucanatide also effectively improved colitis symptoms in TCRa(-/-) mice (a spontaneous colitis model). Because purcanatide exhibited higher anti-proteolytic activity in simulated gastric and intestinal fluids, it was selected for further investigation. This is the first study to report the therapeutic value of a GC-C agonist as a novel oral mucosal active drug candidate for the treatment of inflammatory bowel disease.

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Drug Warning
/Black Box Warning/ Warning: Severe dehydration risk exists in pediatric patients. Trulance is contraindicated in patients under 6 years of age; in a non-clinical study in young mice, a single oral dose of purcanatide resulted in death due to dehydration. Avoid use of Trulance in patients aged 6 to 18 years of age. The safety and efficacy of Trulance in patients under 18 years of age have not been established.
Since the systemic absorption of oral purcanatide and its active metabolites is negligible, it is not expected that administration of this drug to the fetus in pregnant women will result in fetal exposure.
However, current data on the use of purcanatide in pregnant women are insufficient to assess its risk to the fetus.
It is unclear whether purcanatide is excreted into human milk, whether it affects milk production, or whether it has any effect on breastfed infants. Systemic absorption of purcanatide and its active metabolites via oral administration is negligible. It is unclear whether the negligible systemic absorption observed in adults results in clinically significant drug exposure in breastfed infants. The benefits of breastfeeding and the importance of purcanatide to the mother, as well as the potential adverse effects of the drug or the mother's medical conditions on the breastfed infant, should be weighed. Infant exposure to purcanatide may result in serious adverse reactions. Trulance is contraindicated in patients under 6 years of age due to the risk of severe dehydration. Patients with known or suspected mechanical gastrointestinal obstruction. For more complete data on drug warnings (of 10) for purcanatide, please visit the HSDB record page. Pharmacodynamics: Food effects: Subjects who consumed a low-fat, low-calorie (LF-LC) or high-fat, high-calorie (HF-HC) meal within 24 hours after a single 9 mg dose (3 times the recommended dose) reported looser stools than fasting subjects. In clinical studies, purcanatide can be taken with or without food. Prucanatide is a structural analogue of guanosine, differing from it only in one amino acid (Asp³ replaced by Glu³). [1] It is considered a locally acting oral medication for the treatment of inflammatory bowel disease and has the potential to restore intestinal barrier function and reduce inflammation through the GC-C/cGMP signaling pathway. [1] Studies have shown that purcanatide may help delay the progression of colitis to colorectal cancer. [1]

C65H104N18O26S4

1681.89

1680.625

C, 46.42; H, 6.23; N, 14.99; O, 24.73; S, 7.62

467426-54-6

Plecanatide acetate; 1075732-84-1

70693500

White to off-white solid powder

1.5±0.1 g/cm3

2120.0±65.0 °C at 760 mmHg

1235.9±34.3 °C

0.0±0.6 mmHg at 25°C

1.631

-4.45

23

31

28

113

3490

16

S1C[C@@]2([H])C(N[C@H](C(N[C@@H](CC(N)=O)C(N[C@H](C(N[C@@H](C)C(N[C@]([H])(C(N[C@]([H])(C(NCC(N[C@H](C(N[C@H](C(=O)O)CC(C)C)=O)CS1)=O)=O)[C@@H](C)O)=O)CSSC[C@@H](C(N[C@@H](CCC(=O)O)C(N[C@H](C(N2)=O)CC(C)C)=O)=O)NC([C@H](CCC(=O)O)NC([C@H](CC(=O)O)NC([C@H](CC(N)=O)N)=O)=O)=O)=O)=O)C(C)C)=O)=O)C(C)C)=O

NSPHQWLKCGGCQR-DLJDZFDSSA-N

InChI=1S/C65H104N18O26S4/c1-25(2)15-34-55(98)80-41-24-113-110-21-38(58(101)77-37(65(108)109)16-26(3)4)71-44(87)20-69-62(105)50(30(10)84)83-61(104)40(78-51(94)29(9)70-63(106)48(27(5)6)81-57(100)35(18-43(68)86)76-64(107)49(28(7)8)82-60(41)103)23-112-111-22-39(59(102)73-32(53(96)75-34)11-13-45(88)89)79-54(97)33(12-14-46(90)91)72-56(99)36(19-47(92)93)74-52(95)31(66)17-42(67)85/h25-41,48-50,84H,11-24,66H2,1-10H3,(H2,67,85)(H2,68,86)(H,69,105)(H,70,106)(H,71,87)(H,72,99)(H,73,102)(H,74,95)(H,75,96)(H,76,107)(H,77,101)(H,78,94)(H,79,97)(H,80,98)(H,81,100)(H,82,103)(H,83,104)(H,88,89)(H,90,91)(H,92,93)(H,108,109)/t29-,30+,31-,32-,33-,34-,35-,36-,37-,38-,39-,40-,41-,48-,49-,50-/m0/s1

(2S)-2-[[(1R,4S,7S,10S,13S,16R,19S,22S,25R,32S,38R)-10-(2-amino-2-oxoethyl)-25-[[(2S)-4-carboxy-2-[[(2S)-3-carboxy-2-[[(2S)-2,4-diamino-4-oxobutanoyl]amino]propanoyl]amino]butanoyl]amino]-22-(2-carboxyethyl)-32-[(1R)-1-hydroxyethyl]-4-methyl-19-(2-methylpropyl)-3,6,9,12,15,18,21,24,30,33,36-undecaoxo-7,13-di(propan-2-yl)-27,28,40,41-tetrathia-2,5,8,11,14,17,20,23,31,34,37-undecazabicyclo[14.13.13]dotetracontane-38-carbonyl]amino]-4-methylpentanoic acid

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)

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.)