Synthesis of 2-(5-(2-aminobenzylidene)-4-methyl-4,5-dihydrooxazol-2-yl)-3-(2-aminophenyl) acrylonitrile

Abstract

The synthesis and optimization of 2-(5-(2-aminobenzylidene)-4-methyl-4,5-dihydrooxazol-2-yl)-3-(2-aminophenyl) acrylonitrile (PS7) was conducted using Central Composite Design (CCD) for efficient exploration of process variables. The Response Surface Methodology (RSM) was applied to optimize yield based on three key factors: temperature (A), solvent (B), and time (C). ANOVA analysis indicated that the quadratic terms of temperature and solvent were statistically significant, contributing to the overall yield. With a model F-value of 4.55 (p-value: 0.0333), the process was considered significant, suggesting a low probability of error. The model explained 80.36% of the variability in yield (R² = 0.8036), but a predicted R² of -0.4825 highlighted a slight lack of predictive accuracy. Regression equations derived from coded and actual factors allow for yield predictions at specific parameter levels, offering insight into the influence of each factor on the synthesis process. Further model reductions were suggested to improve predictive capabilities. The two-step synthesis involved combining malononitrile, 2-aminopropan-1-ol, acetic acid, and ethanol, followed by refluxing and purification, yielding 87%. In the second step, the intermediate was reacted with 2-aminobenzaldehyde to produce PS7 with an 82% yield and a melting point of 197-199°C. Structural characterization using IR, ¹H-NMR, ¹³C-NMR, and mass spectrometry confirmed the presence of NH, aromatic CH, C=N, C=C, and aliphatic groups. The molecular ion peak at m/z 330 and analytical data matched the expected values for C20H18N4O. The optimized process provides an efficient route for PS7 synthesis with potential for further applications.

Specification

Description:As used herein, 'Amino' represents -NH2.
As used herein, -NH2 group refers to substituent's at 2-(5-(2-aminobenzylidene)-4-methyl-4,5-dihydrooxazol-2-yl)-3-(2-aminophenyl) acrylonitrile ring of Formula I. Exemplary these substituent's include, substitutions at 2nd position of 2-(5-(2-aminobenzylidene)-4-methyl-4,5-dihydrooxazol-2-yl)-3-(2-aminophenyl) acrylonitrile ring of Formula I.








As used herein and in the appended claims, the singular forms "a", "an", and "the" include plural reference unless the context clearly indicates otherwise.
As used herein, the term 'subject' or 'patient' means mammals, such as humans and other animals, including horses, dogs, cats, rats, mice, sheep, pigs, monkeys, chimpanzees or other apes or primates. In exemplary embodiments, the subject may include subjects for which treatment and/or prevention of the conditions described herein would be beneficial.
For ease of reference, in this application, it will be described in terms of administration to human subjects. It will be understood, however, that such descriptions are not limited to, administration to humans, but will also include administration to other animals unless explicitly stated otherwise.
A 'therapeutically effective amount' is the amount of a compound of the present application that is effective in generating biological or medical response of a subject, for example, reduction or inhibition of an enzyme or a protein activity, or ameliorate symptoms, alleviate conditions, slow or delay disease progression, or prevent a disease.
In one embodiment, the term 'a therapeutically effective amount' refers to the amount of the compound of the present application that, when administered to a subject, is effective in (i) at least partially alleviating, inhibiting, preventing and/or ameliorating a condition, or a disorder or a disease mediated by cancer activity.
In another embodiment, the term "a therapeutically effective amount" refers to the amount of the compound of the present invention that, when administered to a cell, or a tissue, or a non-cellular biological material, or a medium, is effective to at least partially reducing or inhibiting the cancer activity.
The terms 'treating' or 'to treat' means to alleviate symptoms, eliminate the causation either on a temporary or permanent basis, or to prevent or slow the appearance of symptoms. The term 'treatment' includes alleviation, elimination of causation of or prevention of any of the diseases or disorders described above. The compound described herein are typically administered in admixture with one or more pharmaceutically acceptable excipients or carriers in the form of a pharmaceutical composition. A 'composition' may contain one compound or a mixture of compound. A 'pharmaceutical composition' is any composition useful in producing at least one physiological response in a subject to which such pharmaceutical composition is administered.
The term 'substantially pure' means that the isolated material is at least 80% pure, preferably 90% pure, more preferably 95% pure, and even more preferably 99% pure as measured by a suitable analytical technique known in the art.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art.
One or more compound of formula (I) can be supplied in the form of a therapeutic composition that is within the scope of the present application.
The term 'Pharmaceutically acceptable salts' refers to any acid or base salt, pharmaceutically acceptable solvates, or any complex of the compound that, when administered to a recipient, is capable of providing (directly or indirectly) a compound as described herein. It should be appreciated, however, that salts that are not pharmaceutically acceptable also lie within the scope of the application. The preparation of salts can be carried out using known methods.
For example, pharmaceutically acceptable salts of compound of formula (I) contemplated refers to salts prepared from acids or bases, including inorganic or organic acids and inorganic or organic bases by conventional chemical methods using a compound of formula (I). Generally, such salts may be prepared, for example, by making free base of the compound and reacting with a stoichiometric quantity of the appropriate acid and vice-versa in water or in an organic solvent, or in a mixture of the two. The compound of the present applications may form mono, di or tris salts.
When the compound of formula (I) is basic, salts may be prepared from acids, including inorganic or organic acids (acid addition salts). Examples of such acids include, but not limited toformic, acetic, trifluoroacetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, mesylic, stearic, salicylic, p-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic),nitric, hydrochloride, hydrobromide, isoethionic, hydroiodide, phosphoric, sulfuric, succinic, tartaric, methanesulfonic, ethanesulfonic, benzenesulfonic, benzoic, mucic, pantothenic, p-toluenesulfonic, camphorsulfonic, 2-hydroxyethanesulfonic, sulfanilic, cyclohexylaminosulfonic, algenic, β-hydroxybutyric, galactaric, and galacturonic acid, and the like.
Salts formed from inorganic bases include sodium, potassium, lithium, calcium, copper, magnesium, manganic salts, manganous, zinc, aluminum, ammonium, ferric, ferrous and the like.
Salts derived from organic bases include salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N,N-dibenzylethylene-diaminesdiethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylene diamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrab amine, isopropylamine, lysine, methylgiucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like. 'Pharmaceutically acceptable salts' in the solid form may exist in more than one crystal structure, and may also be in the form of hydrates.
The term 'stereoisomers' is a general term used for all isomers of an individual molecule that differ only in the orientation of their atoms in space, where the compound according to the present application possess one or more asymmetric centers and compound with asymmetric centers give rise to enantiomers, diastereomersor both as pure or partially purified compound. It is to be understood that all stereoisomeric forms of the compound of the invention, including but not limited to, diastereomers, enantiomers and atropsiomers, as well as mixtures thereof such as forms, are included in the scope of the present application.
(I)
Pharmaceutically acceptable solvates of compound of formula (I) may be hydrates or comprising other solvents of crystallization such as alcohols. Pharmaceutically acceptable solvates of compound of formula (I) may be prepared by conventional methods such as dissolving the compound of formula (I) in solvents such as water, methanol, ethanol etc., preferably water and recrystallizing by using different crystallization techniques.
In the formulae depicted herein, a bond to a substituent and/or a bond that links a molecular fragment to the remainder of a compound may be shown as intersecting one or more bonds in a ring structure. This indicates that the bond may be attached to any one of the atoms that constitutes the ring structure, so long as a hydrogen atom could otherwise be present at that atom. Where no particular substituent(s) is identified for a particular position in a structure, then hydrogen(s) is present at that position.
Reference will now be made in detail to the embodiments of the invention, one or more examples of which are set forth below. Each example is provided by way of explanation of the invention, and not by way of limitation of the invention. In fact, it will be apparent to those skilled in the art that various modification and variations can be made in the present invention without departing from the scope or spirit of the invention. For instance, features illustrated or described as part of one embodiment can be used on another embodiment to yield a still further embodiment. Thus, it is intended that the present invention cover such modifications and variations as come within the scope of the appended claims and their equivalents. Other objects, features, and aspects of the present invention are disclosed in, or are obvious from, the following detailed description. It is to be understood by one of ordinary skill in the art that the present discussion is a description of exemplary embodiments only, and is not to be construed as limiting the broader aspects of the present application.
Thus, in accordance of this application there is provided a 2-(5-(2-aminobenzylidene)-4-methyl-4,5-dihydrooxazol-2-yl)-3-(2-aminophenyl) acrylonitrile compound having the general formula (I),
The present applications relate to 2-(5-(2-aminobenzylidene)-4-methyl-4,5-dihydrooxazol-2-yl)-3-(2-aminophenyl) acrylonitrile compound of formula (I),
(I)
and pharmaceutically acceptable salts, solvates or stereoisomers thereof;
Wherein said R is - NH2 in 2nd position in Formula I

or pharmaceutically acceptable salts thereof, pharmaceutically acceptable solvates thereof or stereoisomers thereof; wherein
In another embodiment, the compound of formula (I) is represented as compound of formula (Ia1),
In an embodiment, the present application relates to a process of preparing 2-(5-(2-aminobenzylidene)-4-methyl-4,5-dihydrooxazol-2-yl)-3-(2-aminophenyl)acrylonitrile compound of formula (I) comprising steps of:
Synthesis
Optimization of synthesis using CCD
The RSM analysis for yield optimization of any PS7 synthesis process. The ANOVA (Analysis of Variance) tables and regression equations for a quadratic model with three factors, Temperature (A), Solvent(B) or Time(C). From the model, it is seen that temperature and solvent time quadratic are statistically significant while as per equation also implying, they have an effect on yield. With the model f-value of 4.55, thus can say that this is a significant process (p value:0,0333) and no matter how many times we repeat testing only very less will be by chance of probability for error. While an R² of 0.8036 means that 80.36% variability in the yield have been explained by our model, but a predicted R² -0.4825 shows slight lack of predictive ability. The final equations obtained based on coded and actual factors can be employed to predict the yield at particular factor levels, for only understanding the relevance weightage of each factor requires considering them as per coded values. Some recommendations for model reduction to enhance prediction accuracy.
Formula: Step 1
Yield=−2016.59843+32.25391⋅Temperature+34.57526⋅Solvent+2.87459⋅Time+7.26494×10−15⋅ (Temperature×Solvent)+1.42018×10−15⋅(Temperature×Time)+3.47722×10−15⋅(Solvent×Time)−0.213917⋅Temperature2−0.383622⋅Solvent2−0.019054⋅Time2
Formula: Step 2
Yield=−2760.22044+7.96730⋅Temperature+7.07298⋅Solvent+41.21543⋅Time+6.08529×10−15⋅ (Temperature×Solvent)+5.61929×10−15⋅(Temperature×Time)+1.86455×10−14⋅(Solvent×Time)−0.037196⋅ Temperature2−0.235766⋅Solvent2−0.179198⋅Time2
To optimize synthesis using the CCD (Central Composite Design) in Design Expert software, first define the factors and response variables relevant to your synthesis. Set the desired range for each factor and use the CCD module to generate an experimental design that efficiently explores variable interactions. After running the experiments, input the data into Design Expert for analysis. The software helps identify optimal conditions by generating response surface plots, contour plots, and predictive models. Adjustments to synthesis parameters are suggested based on statistical significance and desirability functions.
Step 1: Combine malononitrile (50 mmol), 2-aminopropan-1-ol (50 mmol), acetic acid (50 mL), and ethanol (20 mL) in a flask. Heat the mixture at reflux with stirring for 60 minutes at 70°C. Let it cool for 30 minutes, filter the precipitate, wash with water, recrystallize from alcohol, and dry (yield: 87%). Step 2: Dissolve 1I (2 mmol) in ethanol (8.5 mL), add 2-aminobenzaldehyde, and reflux for 115 minutes at 110°C. Cool the mixture, filter the precipitate, wash with cold ethanol, recrystallize from boiling ethanol, and dry (yield: 82%).
2-(5-(2-aminobenzylidene)-4-methyl-4,5-dihydrooxazol-2-yl)-3-(2-aminophenyl) acrylonitrile PS7
Yield: 82%; m.p.: 197-199 °C. IR (cm-1): 3281 (NH), 3017 (Ar-CH), 2950 (Ali-CH), 1661 (C=N), 1626 (C=C), 1013 (C-O-C).
1H-NMR (δ: ppm): 6.91-7.72 (11H, m, Ar-CH), 3.94 (2H, NH2), 2.93 (1H, s, Aliphatic-CH), 1.69 (3H, CH3).
13C-NMR (δ: ppm): 176.4, 153.4 (2C, C=C), 143.4 (1C, Carbonyl Carbon (C-NH2)), 115.6-132.1 (13C, Aromatic Carbon), 57.2 (1C, Alicyclic Carbon), and 28.6 (1C, Aliphatic Carbon). MS (EI) m/z: .330 (M+). Anal. Calcd for C20H18N4O: C, 76.17; H, 5.43; N, 13.32
The compound 2-(5-(2-aminobenzylidene)-4-methyl-4,5-dihydrooxazol-2-yl)-3-(2-aminophenyl) acrylonitrile (PS7) was synthesized with an 82% yield and a melting point of 197-199 °C. Its IR spectrum shows characteristic peaks for NH (3281 cm⁻¹), aromatic CH (3017 cm⁻¹), aliphatic CH (2950 cm⁻¹), C=N (1661 cm⁻¹), and C=C (1626 cm⁻¹). ¹H-NMR and ¹³C-NMR data confirm the presence of aromatic, aliphatic, and amine groups. The molecular ion peak in the mass spectrum appears at m/z 449. The analytical data for carbon, hydrogen, and nitrogen content are consistent with the calculated values for the compound's molecular formula C20H18N4O.

The provided procedures here are a part of invention and are not restricting the scope of invention in any manner.
The performed pharmacophore mapping was used to study the structural features of the synthesized compound to identify the vital moieties necessary for effective and selective cancer activities.
Scheme

, Claims:1.A pharmaceutical compound of formula (I)
pharmaceutically acceptable salts, solvates or stereoisomers thereof;
wherein R is '-NH2,
The compound of claim 1, wherein said compound is in substitution and dependent
2. A method of treating a breast cancer patient comprising: administering a therapeutically effective amount of formula (I) to a patient in need thereof.
3. The method of claim 3, wherein said compound is administered orally, topically, parenterally and a combination thereof

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