Actin polymerization

Cytochalasin D (3 and 10 μM; 30 min) induces F-actin to change from long fibers to punctate structures, as well as the contraction and branching of COS-7 cells [1]. In COS-7 cells, cytochalasin D (0.3, 1, 3, and 10 μM; 30 min) significantly lowers the rate of actin depolymerization [1]. Yap phosphorylation and fibrosis are both disrupted when cytokinin D (1 μM) is shortened in NIH3T3 cells, although the cells maintain their previous state.

The antitumor effects of cytochalasin B were mirrored by cytochalasin D at much lower concentrations (Fig. 6b). It only took 2 mg/kg/day cytochalasin D administered for eight consecutive days (Days 1–8) to produce marked prolongation in the life expectancy of mice challenged with P388/S, as well as a 20 % long-term survival rate. Whether or not cytochalasin D would exhibit the same antitumor effect against P388/ADR at these lower concentrations remains unclear, as not enough mice remained to establish another treatment group of sufficient quantity. Nevertheless, it is likely that at least some prolongation in life expectancy would be observed. The cytochalasin vehicle CMC/Tw did not affect the life span of mice challenged with either leukemia, demonstrating that there is no effect of the lipophilic detergent vehicle on the leukemia challenges in the absence of cytochalasins.[Invest New Drugs. 2015 Apr;33(2):290-9.]

Cofilin, a key regulator of actin filament dynamics, binds to G- and F-actin and promotes actin filament turnover by stimulating depolymerization and severance of actin filaments. In this study, cytochalasin D (CytoD), a widely used inhibitor of actin dynamics, was found to act as an inhibitor of the G-actin-cofilin interaction by binding to G-actin. CytoD also inhibited the binding of cofilin to F-actin and decreased the rate of both actin polymerization and depolymerization in living cells. CytoD altered cellular F-actin organization but did not induce net actin polymerization or depolymerization. These results suggest that CytoD inhibits actin filament dynamics in cells via multiple mechanisms, including the well-known barbed-end capping mechanism and as shown in this study, the inhibition of G- and F-actin binding to cofilin[1].

Western Blot Analysis
Cell Types: COS-7 cells[1]
Tested Concentrations: 3 and 10 μM
Incubation Duration: 30 min
Experimental Results: Causes the retraction and arborization of COS-7 cells and the transformation of F-. Actin changes from long fibers to punctate structures.

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Western Blot Analysis
Cell Types: COS-7 cells expressing YFP-actin [1]
Tested Concentrations: 0.3, 1, 3 and 10 μM
Incubation Duration: 30 minutes
Experimental Results: Concentration-dependent decrease in actin lysis in COS-7 cells polymerization rate.

P388 leukemias in vivo[Invest New Drugs. 2015 Apr;33(2):290-9.]
For chemotherapy testing, Balb/c mice under isoflurane anesthesia were challenged with 2 × 105 trypan blue negative P388/S or P388/ADR cells subcutaneously (s.c.) in a volume of 200 μl. Untreated mice were kept in order to determine the lethality of the challenge without chemotherapeutic intervention. Long-term survival was defined as challenged mice that survived the duration of the observation period.

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Cytochalasin D intraperitoneal administration[Invest New Drugs. 2015 Apr;33(2):290-9.]
Cytochalasin D was prepared in suspension form in 2 % carboxymethyl cellulose 1 % tween 20 (CMC/Tw) for intraperitoneal (i.p.) administration, as previously described. The congeners or the vehicle were administered to leukemia-challenged mice on Days 1–8 following the initial challenge.

Metabolism / Metabolites
IN...WORK USING STEREOSPECIFICALLY LABELLED PRECURSORS, L-PHENYLALANINE WAS SHOWN TO BE THE PRIMARY PRECURSOR OF CYTOCHALASIN D, & THE RAPID EQUILIBRIUM OF D- & L-ISOMERS THROUGH PHENYLPYRUVIC ACID WAS SHOWN TO ACCOUNT FOR THE EQUALLY EFFICIENT INCORPORATION OF BOTH ENANTIOMERS.
IN...THE BIOSYNTHESIS OF CYTOCHALASIN D BY ZYGOSPORIUM MASONII, [1,2-(13)C]-ACETATE WAS FED TO A GROWING CULTURE OF THIS FUNGUS. THE ARRANGEMENT OF INTACT ACETATE UNITS WAS ESTABLISHED... CYTOCHALASIN D ORIGINATED FROM PHENYLALANINE, METHIONINE & 9 INTACT ACETATE UNITS. EIGHT OF THE ACETATE UNITS COUPLE IN A HEAD-TO-TAIL FASHION TO FORM THE C16-POLYKETIDE MOIETY. THE FEEDING OF HIGHER EVEN-NUMBERED SATURATED ACIDS SUCH AS C1-LABELLED BUTYRATE, MYRISTATE & PALMITATE SHOWED NO DIRECT INCORPORATION. THEY DO, HOWEVER, UNDERGO BETA-OXIDATION TO YIELD C1-LABELLED ACETATE WHICH IS INCORPORATED INTO CYTOCHALASIN D.

Rat LD50 intraperitoneal 900 ug/kg

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Interactions
PRETREATMENT OF RABBIT KIDNEY CELLS WITH CYTOCHALASIN D ENHANCED HERPES SIMPLEX VIRUS TYPE 2 INFECTIVITY 3- TO 6-FOLD OVER VALUES OBTAINED USING THE STANDARD CALCIUM CHLORIDE TECHNIQUE.
CYTOCHALASIN D TREATMENT OF PERMISSIVE OR SEMIRESTRICTIVE CELL LINES ENHANCED THEIR SUSCEPTIBILITY TO INFECTION WITH POLIOVIRUS.

[1]. Cytochalasin D acts as an inhibitor of the actin-cofilin interaction. Biochem Biophys Res Commun. 2012 Jul 20;424(1):52-7.

[2]. Cytochalasins block actin filament elongation by binding to high affinity sites associated with F-actin. J Biol Chem. 1980 Feb 10;255(3):835-8.

[3]. Inhibiting extracellular vesicles formation and release: a review of EV inhibitors. J Extracell Vesicles. 2019 Dec 19;9(1):1703244.

[4]. Hippo pathway regulation by cell morphology and stress fibers. Development. 2011 Sep;138(18):3907-14.

Cytochalasin d appears as needles or fluffy white powder. (NTP, 1992)
cytochalasin D has been reported in Ascochyta rabiei, Zygosporium masonii, and Metarhizium anisopliae with data available.
A fungal metabolite that blocks cytoplasmic cleavage by blocking formation of contractile microfilament structures resulting in multinucleated cell formation, reversible inhibition of cell movement, and the induction of cellular extrusion. Additional reported effects include the inhibition of actin polymerization, DNA synthesis, sperm motility, glucose transport, thyroid secretion, and growth hormone release.

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Mechanism of Action
MAJOR BIOLOGICAL EFFECTS ARE THE BLOCKAGE OF CYTOPLASMIC CLEAVAGE RESULTING IN MULTINUCLEATE CELL FORMATION, THE INHIBITION OF CELL MOVEMENT, & THE INDUCTION OF NUCLEAR EXTRUSION... OTHER REPORTED EFFECTS INCLUDE THE INHIBITION OF GLUCOSE TRANSPORT, OF THYROID SECRETION, OF GROWTH HORMONE RELEASE, OF PHAGOCYTOSIS, & OF PLATELET AGGREGATION & CLOT CONTRACTION. /CYTOCHALASINS/
IN THE EARLY STAGE OF CYTOLOGICAL RESEARCH USING CYTOCHALASINS THE MODE OF ACTION OF THE COMPOUNDS WAS EXPLAINED AS DIRECT DISRUPTION OF MICROFILAMENTS INTERFERRING WITH CONTRACTILE MOVEMENTS OF THE MAMMALIAN CELLS. FURTHER STUDY, ESPECIALLY THE OBSERVATION OF THEIR EFFECTS ON THE TRANSPORT OF A NUMBER OF SUBSTANCES INTO THE CELLS, REVEALED A MORE COMPLEX MODE OF ACTION. IT IS CLEAR THAT THE NATURE OF THE BINDING BETWEEN THE CYTOCHALASINS & ACTIVE SITES IS NOT COVALENT BECAUSE OF THE RAPID REVERSIBILITY OF THE ACTION. /CYTOCHALASINS/
THROUGH THE USE OF (3)H-LABELLED CYTOCHALASINS B & D THE PRECISE BINDING SITES & SUB-CELLULAR LOCALIZATION ARE NOW UNDER INVESTIGATION... EXPERIMENTAL RESULTS OBTAINED THUS FAR SUPPORT THE IDEA THAT THE PRIMARY EFFECT IS MEMBRANOTROPIC, PERHAPS INVOLVING THE ASSOCIATION OF MICROFILAMENTS WITH THE PLASMA MEMBRANE.
CYTOCHALASIN D DID NOT PREVENT ATTACHMENT BUT DID PREVENT ENTRY OF TOXOPLASMA GONDII INTO PERITONEAL MACROPHAGES AND BLADDER TUMOR 4934 CELLS.
For more Mechanism of Action (Complete) data for CYTOCHALASIN D (9 total), please visit the HSDB record page.

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Therapeutic Uses
Nucleic Acid Synthesis Inhibitors
EXPTL USE: ...WORKERS INVESTIGATED THE APPLICATION OF.../ZYGOSPORIN A (CYTOCHALASIN D), E, F, G, & MORE THAN 30 DERIVATIVES/ AS ANTITUMOR AGENTS BUT WITH UNSUCCESSFUL RESULTS.
EXPTL USE: CYTOCHALASIN D (0.322-5.0 MG/KG/DAY, IP) HAD ANTITUMOR ACTIVITY AGAINST ASCITES HEPATOMA AH-130 AND MURPHY-STURM LYMPHOSARCOMA IN RATS.

C30H37NO6

507.62

507.262

C, 70.98; H, 7.35; N, 2.76; O, 18.91

22144-77-0

5458428

White to off-white solid powder

1.2±0.1 g/cm3

712.1±60.0 °C at 760 mmHg

255-260ºC

384.5±32.9 °C

0.0±2.4 mmHg at 25°C

1.597

2.64

3

6

4

37

996

9

C[C@H]1C/C=C/[C@H]2[C@@H](C(=C)[C@H]([C@@H]3[C@@]2([C@@H](/C=C/[C@@](C1=O)(C)O)OC(=O)C)C(=O)N[C@H]3CC4=CC=CC=C4)C)O

SDZRWUKZFQQKKV-JHADDHBZSA-N

InChI=1S/C30H37NO6/c1-17-10-9-13-22-26(33)19(3)18(2)25-23(16-21-11-7-6-8-12-21)31-28(35)30(22,25)24(37-20(4)32)14-15-29(5,36)27(17)34/h6-9,11-15,17-18,22-26,33,36H,3,10,16H2,1-2,4-5H3,(H,31,35)/b13-9+,15-14+/t17-,18+,22-,23-,24+,25-,26+,29+,30+/m0/s1

[(1R,2R,3E,5R,7S,9E,11R,12S,14S,15R,16S)-16-benzyl-5,12-dihydroxy-5,7,14-trimethyl-13-methylidene-6,18-dioxo-17-azatricyclo[9.7.0.01,15]octadeca-3,9-dien-2-yl] acetate

Lygosporin A; Zygosporin A; 22144-77-0; Zygosporin A; Cytohalasin D; Lygosporin A; MFCD00077706; SY9F0FZ3TO; NSC209835; Cytochalasin D

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)

DMSO : ~25 mg/mL (~49.25 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.)