Plasminogen Activator Inhibitor-1 (PAI-1) mRNA (induction) [1]

Anecortave Acetate is designed to be devoid of conventional hormonal activity. It contains a 9-11 double bond replacing the 11β-hydroxyl group, which is essential for glucocorticoid and mineralocorticoid activities. It demonstrates no significant glucocorticoid-mediated anti-inflammatory agonist activity in in vitro inflammation assays, including IL-1 induction in cultured human U937 cells. [1]
Anecortave Acetate does not block the anti-inflammatory activity of dexamethasone, indicating it is devoid of glucocorticoid antagonist activity. [1]

Anecorstat acetate, when administered intraocularly (5 μl 10% solution), markedly reduces pathological retinal angiogenesis and raises levels of PAI-1 mRNA [2].

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In a rat model of Retinopathy of Prematurity (ROP), a single intravitreal injection of 5 μl of a 10% suspension of Anecortave Acetate significantly reduced the severity of abnormal retinal neovascularization compared to vehicle-injected eyes. The median neovascularization score was 2 clock hours (range 0-4) for drug-treated eyes versus 6 clock hours (range 3-8) for vehicle-treated eyes when injected on Day 14 (P < 0.0001). For injections on Day 16, the median score was 3 clock hours (range 1-5) for drug-treated eyes versus 6 clock hours (range 3-10) for vehicle-treated eyes (P = 0.002). [1]
The extent of normal total retinal vascular area was not significantly different between Anecortave Acetate-treated and vehicle-treated eyes in the ROP rat model, although a trend toward slight inhibition (approximately 15%) was observed. In room air-raised control rats, the drug had no significant effect on retinal vascular area or neovascularization. [1]
Eyes injected with Anecortave Acetate in the ROP rat model demonstrated a six- to ninefold increase in PAI-1 mRNA levels at 1 to 3 days after injection, as measured by RNase protection assay. This induction was rapid (by 24 hours), sustained (at least 3 days), and robust. [1]
In rabbit corneal neovascularization models, topical application of Anecortave Acetate resulted in 90% inhibition of the area of new corneal blood vessels. [1]
In an intraocular tumor model, topical application of Anecortave Acetate held net tumor weight to less than one-third that of the control, an effect concluded to be the result of its angiostatic properties. [1]

Animal/Disease Models: SD (SD (Sprague-Dawley)) albino rat (intravitreal injection of premixed antibiotics to affect retinal blood vessel growth) [2]
Doses: 5 μl 10% anectaline acetate suspension
Route of Administration: Injection into the eye
Experimental Results: Significant inhibition of the Pathological retinal angiogenesis in the model without Dramatically affecting normal intraretinal blood vessels. PAI-1 mRNA increased 6- to 9-fold 1 to 3 days after injection.

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To test the effect of Anecortave Acetate on retinal neovascularization, newborn Sprague-Dawley rats were placed in a variable oxygen environment (alternating 50% and 10% every 24 hours) for 14 days to induce ROP. At 14 days of age (immediately after removal from the oxygen chamber), rats received a single intravitreal injection in the left eye. Half received 5 μl of a 10% suspension of Anecortave Acetate, and the other half received 5 μl of vehicle (70:20:10 by volume, polyethylene glycol: phosphate-buffered saline: EtOH). This injection protocol was repeated in the right eye of each animal at 16 days of age. All rats were subsequently kept in room air and were euthanized at 18 days of age for retinal analysis. [1]
* To determine the effect of Anecortave Acetate on normal retinal vascular development, 7-day-old rats raised in room air received an intravitreal injection of 5 μl of a 10% suspension of the drug in the left eye and 5 μl of vehicle in the right eye. Control rats received no injection. These rats were euthanized at 10 days of age for retinal analysis. [1]
* For the RNase protection assay mechanism study, 14-day-old rats from the variable oxygen environment were given an intravitreal injection of Anecortave Acetate, vehicle, or nothing. They were then euthanized 1, 2, or 3 days later, and their retinas were dissected for mRNA analysis. [1]

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To test the effect of Anecortave Acetate on retinal neovascularization, newborn Sprague-Dawley rats were placed in a variable oxygen environment (alternating 50% and 10% every 24 hours) for 14 days to induce ROP. At 14 days of age (immediately after removal from the oxygen chamber), rats received a single intravitreal injection in the left eye. Half received 5 μl of a 10% suspension of Anecortave Acetate, and the other half received 5 μl of vehicle (70:20:10 by volume, polyethylene glycol: phosphate-buffered saline: EtOH). This injection protocol was repeated in the right eye of each animal at 16 days of age. All rats were subsequently kept in room air and were euthanized at 18 days of age for retinal analysis. [1]
To determine the effect of Anecortave Acetate on normal retinal vascular development, 7-day-old rats raised in room air received an intravitreal injection of 5 μl of a 10% suspension of the drug in the left eye and 5 μl of vehicle in the right eye. Control rats received no injection. These rats were euthanized at 10 days of age for retinal analysis. [1]
For the RNase protection assay mechanism study, 14-day-old rats from the variable oxygen environment were given an intravitreal injection of Anecortave Acetate, vehicle, or nothing. They were then euthanized 1, 2, or 3 days later, and their retinas were dissected for mRNA analysis. [1]

The ocular pharmacokinetics of Anecortave Acetate are only partly defined, but daily examination of injected eyes in this study suggested that the highly hydrophobic compound was not rapidly cleared from the vitreous. [1]

Anecortave Acetate is described as being relatively nontoxic, with no studies describing its use reporting attrition. [1]
It is designed to be devoid of conventional hormonal (glucocorticoid and mineralocorticoid) activity, which is important because of the significant ocular side effects associated with glucocorticoid therapy. [1]

[1]. Clark AF. Mechanism of action of the angiostatic cortisene anecortave acetate. Surv Ophthalmol. 2007 Jan;52 Suppl 1:S26-34.

[2]. The effect of an angiostatic steroid on neovascularization in a rat model of retinopathy of prematurity. Invest Ophthalmol Vis Sci. 2001 Jan;42(1):283-90.

Anecortave acetate is an organic molecular entity. Anecortave acetate (Retaane) is an analogue of corticosteroid acetate; its steroid modifications include the removal of the 11β-hydroxyl group (OH) and the addition of a 21-acetic acid ester group. Due to these modifications, Anecortave acetate lacks the typical anti-inflammatory and immunosuppressive properties of glucocorticoids. Alcon is developing and marketing Retaane. See also: Anecortave (note moved to).

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Drug Indications
It has been studied for the treatment of glaucoma and macular degeneration.

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Mechanism of Action
Anecortave acetate acts as an anti-angiogenic agent, inhibiting angiogenesis by reducing the expression of extracellular proteases and inhibiting endothelial cell migration. Its anti-angiogenic activity does not appear to be mediated through any known common pharmacological receptors. (Ophthalmology 2004;111:2316-7) RETAANE blocks the signaling of multiple growth factors because it acts downstream, independently of initiating angiogenesis stimulation, and inhibits angiogenesis after angiogenesis stimulation.

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Anecortave Acetate [4,9(11)-Pregnadien-17a,21-diol-3,20-dione-21-acetate] is an angiostatic steroid. Its chemical structure includes a 9-11 double bond that replaces the 11β-hydroxyl group, a modification that eliminates traditional glucocorticoid and mineralocorticoid activities. [1]
The proposed mechanism of action for Anecortave Acetate is the induction of Plasminogen Activator Inhibitor-1 (PAI-1). Increased PAI-1 inhibits urokinase-type plasminogen activator (u-PA) activity, which is essential for the breakdown of vascular endothelium basement membrane and extracellular matrix during angiogenesis. This inhibition prevents endothelial cells from proliferating and migrating, thereby suppressing new blood vessel formation. This study provides the first in vivo evidence for this mechanism. [1]
Anecortave Acetate has demonstrated angiostatic capacity in various models, including the chick chorioallantoic membrane, rabbit cornea, and intraocular tumor models. Its bioactivity appears independent of species or the cause of angiogenesis. [1]
The therapeutic effect of Anecortave Acetate in the ROP model is particularly compelling because it profoundly inhibited pathologic neovascularization (≥50% inhibition vs. vehicle) while having a limited effect on normal retinal vessel development (approximately 15% inhibition vs. vehicle). This discrimination may be due to the different processes of vasculogenesis (normal development) and angiogenesis (pathologic growth) or the drug's availability to preretinal vessels. [1]

386.209

7753-60-8

7753-60-8

111332

White to off-white solid powder

1.23g/cm3

551.8ºC at 760 mmHg

188.4ºC

1.572

3.301

1

5

4

28

808

5

CC(OCC([C@@]1([C@]2(CC=C3[C@]4(C)C(=CC(CC4)=O)CC[C@H]3[C@@H]2CC1)C)O)=O)=O

YUWPMEXLKGOSBF-GACAOOTBSA-N

InChI=1S/C23H30O5/c1-14(24)28-13-20(26)23(27)11-8-19-17-5-4-15-12-16(25)6-9-21(15,2)18(17)7-10-22(19,23)3/h7,12,17,19,27H,4-6,8-11,13H2,1-3H3/t17-,19+,21+,22+,23+/m1/s1 SMILES

17,21-Dihydroxypregna-4,9(11)-diene-3,20-dione 21-acetate

Anecortave Acetate Anecortave Al 3789 NSC 15475 NSC 24345 Retaane Hydrocortisone Acetate EP Impurity E

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.)