Inactive form of Colchicine

As an additional control, γ-lumicolchicine, an inactive analog of colchicine, was also used to treat α2β-expressing HEK cells (Fig 4A). These brief treatments had no obvious effect on the survival of untransfected cells. There was a trend for colchicine treatment to reduce the overall current density at 300 μM glycine, 56.7 ± 9.1 pA/pF in control cells (n = 8), 41.2 ± 2.3 pA/pF in colchicine-treated cells (n = 10), and 50.1 ± 9.9 pA/pF (n = 6) in γ-lumicolchicine-treated cells; however, this was not significant (p > 0.05, ANOVA) and was probably not related to any direct action of colchicine given that the glycine current density was also slightly reduced in α2β-expressing cells exposed to γ-lumicolchicine compared to controls. However, the efficacy of both taurine (p < 0.01, One-way ANOVA) and β-alanine (p < 0.05, ANOVA) were significantly decreased by colchicine but not γ-lumicolchicine treatment. Taurine efficacy was 33 ± 6% of glycine in controls, 13 ± 3% following colchicine, and 28 ± 3% following γ-lumicolchicine. Similarly, β-alanine efficacy was 70 ± 7% of glycine in controls, 49 ± 6% following colchicine, and 72 ± 7% following γ-lumicolchicine. Similar treatment of α2-expressing HEK cells with colchicine (Fig. 4B) did not reveal any significant effect on glycine current density (54 ± 14 pA/pF in controls, 60 ± 15 pA/pF in treated), on taurine efficacy (34 ± 16% in controls vs. 38 ± 10% in treated), or on β-alanine efficacy (71 ± 12% in controls vs. 86 ± 8% in treated). Cholchicine treatment also significantly reduced β-alanine efficacy in L-cells expressing GlyRα2β (23 ± 2% in controls vs. 12 ± 3% in treated, P < 0.05, t-test) but not in GlyRα2-expressing L-cells (Fig. 4C). We did not attempt to examine taurine in L-cells treated with colchicine given the exceptionally low efficacy of receptors expressed in this cell line [1].

[1]. Extrinsic factors regulate partial agonist efficacy of strychnine-sensitive glycine receptors. BMC Pharmacol. 2004 Aug 9;4:16.

LSM-4236 is a carbon-tricyclic compound belonging to the acetamide class and is also an alkaloid. β-Photosensitive colchicine has been reported in Colchicum arenarium, Colchicum autumnale, and other organisms with relevant data. The UV degradation products of colchicine have three isomers: α, β, and γ, which lack many of the physiological activities of the parent compound; they can be used as experimental controls for the effects of colchicine. See also: γ-Photosensitive colchicine (note moved here). Background: Many strychnine-sensitive glycine receptors in pre-adult brain regions are composed of α2+β heterologous channels. This subunit composition differs from the α1+β channels prevalent in the adult spinal cord. Unfortunately, the pharmacological properties of forebrain α2β receptors are not well understood compared to neonatal α2 homologous channels or spinal cord α1β heterologous channels. Furthermore, the pharmacological properties of native α2β-glycine receptors often differ from those of receptors produced by heterologous expression. Finding a heterologous expression system highly similar to these native glycine-gated chloride channels is crucial for identifying subtype-specific pharmacological tools for forebrain α2β receptors. Results: In exploring the pharmacological properties of α2β-glycine receptors and α2 homologous channels, we found that different heterologous expression systems appear to have varying effects on the pharmacological properties of partial agonists. When the β-amino acid taurine was expressed in HEK 293 cells, it exhibited 30–50% potency against receptor subtypes containing the α2 subunit. However, the potency of taurine was significantly reduced in L-cell fibroblasts. Similar results were observed with β-alanine. The potency of these partial agonists was also significantly reduced due to the β subunit. Epistylsinic affinity values calculated from concentration-response data did not differ significantly between different expression systems or subunit combinations. No correlation was found between relative expression levels and the potency of partial agonists when comparing within or between several different expression systems. Finally, disruption of the tubulin cytoskeleton reduced the potency of partial agonists in a subunit-dependent but system-independent manner. Conclusion: Our results indicate that different heterologous expression systems can significantly affect the agonist pharmacology of strychnine-sensitive glycine receptors. In the systems studied in this paper, these effects were independent of any system-related alterations in absolute expression levels or agonist binding sites. We conclude that the complex interactions between receptor composition and extrinsic factors may play an important role in determining the partial agonist pharmacology of strychnine-sensitive glycine receptors. [1]

C22H25NO6

399.437

399.168

C, 66.15; H, 6.31; N, 3.51; O, 24.03

6901-14-0

6901-13-9; 6901-14-0

244898

White to off-white solid powder

1.3g/cm3

623.2ºC at 760 mmHg

268ºC

330.7ºC

1.596

2.666

1

6

5

29

758

3

CC(=O)NC1CCC2=CC(=C(C(=C2C3=C1C4C3C=C(C4=O)OC)OC)OC)OC

VKPVZFOUXUQJMW-FHSNZYRGSA-N

InChI=1S/C22H25NO6/c1-10(24)23-13-7-6-11-8-15(27-3)21(28-4)22(29-5)16(11)17-12-9-14(26-2)20(25)18(12)19(13)17/h8-9,12-13,18H,6-7H2,1-5H3,(H,23,24)/t12-,13+,18-/m1/s1

N-[(10S,12R,16S)-3,4,5,14-tetramethoxy-13-oxo-10-tetracyclo[9.5.0.02,7.012,16]hexadeca-1(11),2,4,6,14-pentaenyl]acetamide

gamma-Lumicolchicine; 6901-14-0; EINECS 230-009-7; N-(3,4,5,14-tetramethoxy-13-oxo-10-tetracyclo[9.5.0.02,7.012,16]hexadeca-1(11),2,4,6,14-pentaenyl)acetamide; 490-24-4; Lumicolchicines; gamma Lumicolchicine; N-(3,4,5,14-tetramethoxy-13-oxo-10-tetracyclo(9.5.0.02,7.012,16)hexadeca-1(11),2,4,6,14-pentaenyl)acetamide;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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.)