DESC:Related application
This application claims the benefit of priority of our Indian patent application numbers 3287/MUM/2014 filed on Oct. 14. 2010 and 4214/MUM/2014 filed on Dec. 30, 2014 which are incorporated herein by reference.
FIELD OF THE INVENTION:
The present invention relates to the novel impurities of Bosutinib, preparation of impurities and characterization data thereof which occur due to chemical instability of the target substance and to a process for synthesizing Bosutinib; also, an improved process for the preparation of Bosutinib and its polymorphic forms.
BACKGROUND OF THE INVENTION:
Bosutinib (marketed under the trade name Bosulif) is a tyrosine kinase inhibitor undergoing for use in the treatment of cancer. Originally synthesized by Wyeth, it is being developed by Pfizer. The product marketed form is Bosutinib monohydrate.

Bosutinib is firstly described in US patent RE42376. The patent discloses process for the preparation of Bosutinib generically.
US 6780996 B2 claims process for the preparation of 7-substituted-3-quinolinecarbonitriles.
WO2005019201A2 and CN101792416B also describe a process for the preparation of Bosutinib.
Bosutinib obtained by the processes described in the above prior art does not have satisfactory purity for pharmaceutical use.
In general, the chemical purity of the Active Pharmaceutical Ingredient (API) produced in an industrial scale is one of the critical parameters for its commercialization.
Standards of impurities, analytic methods of chemical purity of the API and quantity determination methods are extremely important for the control of the production process and consequently for successful commercialization of the product.
It is known by those skilled in the art, the management of process impurities is greatly enhanced by understanding their chemical structures and synthetic pathways, and by identifying the parameters that influence the amount of impurities in the final product.
As with drug substance preparations of any synthetic organic API, drug substance preparations of Bosutinib can contain product-related impurities & process-related impurities.
WO2007005462 described form I, form II, form III, form IV, form V and form VI. Form I & II is monohydrated form, form III is mono isopropyl alcoholate form, form IV is likely a hydrated form, form V is anhydrous form and form VI is methanolate form of Bosutinib. This application also mention that Form I is more stable than Form II, III, IV, V and VI. But none of the prior arts discloses amorphous form of Bosutinib. Accordingly, there remains a need in the art to prepare a stable and substantially pure amorphous form of Bosutinib.
SUMMARY OF THE INVENTION:
The invention provides especially analytical standards and analytical methods used for the control of the production process and final quality of Bosutinib.
The invention relates to drug substance contain limited amounts of specific impurities.
The aspect of the present invention is drug substance preparations of Bosutinib having minimal impurities. The invention provides a high yielding, high purity, operationally simple, commercially viable and cost effective process for the preparation of Bosutinib of formula I.
The preparation of Bosutinib comprising,

a. Combination of the compound of formula-VII with cyno acetic acid by using reagents which provides good leaving group to obtain the cyno acetamide compound of formula-VIII;

b. Reacting Compound of Formula III with formula IX in presence of alkali halides and base, with suitable solvent provides compound of formula-IV;

c. Reduction of compound of formula-IV in presence of Pd catalyst and suitable alcholic solvent gives the compound of formula-V;

d. Condensing compound of formula-V with compound of formula-VIII in presence of trialkyl ortho formate, suitable solvent provides compound of formula-VI;

e. Cyclodehydration of compound of formula-VI in presence of phosphorus oxychloride, and suitable solvent provides compound of formula-I;

f. Crystallizing or purifying the crude Bosutinib in presence of suitable solvent system to provide suitable crystalline Form.
Another aspect of the present invention is to provide method of preparation of 2,4-dichloro-5-methoxyaniline from the N-(2,4-dichloro-5-hydroxyphenyl)acetamide having intermediate N-(2,4-dichloro-5-methoxyphenyl)acetamide.

Another aspect, the present invention relates a novel amorphous form of Bosutinib and preparation thereof.
Another aspect, the present invention relates a novel process for preparation of Bosutinib Form-V, comprising using ethanol-methanol mixture and cyclohexane as solvents.
Another aspect of the present invention is to provide novel impurities of Bosutinib such as compound of formula A, formula B, formula C, formula D and formula E.
In another aspect of the invention the novel impurities are used as reference makers.
Another aspect of the present invention is to provide compound of formula A or it’s salt.

Another aspect of the present invention is to provide compound of formula B or its salt.

Another aspect of the present invention is to provide compound of formula C or its salt.

Another aspect of the present invention is to provide compound of formula D or its salt.

Another aspect of the present invention is to provide compound of Formula-E or its salt.

Another aspect of the present invention is to provide a preparation process of compound of formula A, B, C, D, E and to provide characterization data thereof.
Another aspect of the present invention is to provide an isolated specific impurity of Bosutinib, selected from the group including formula A or formula B or formula C or formula D or formula E and their salts, characterized by chemical purity of more than 50% for use in setting the analytic methods designed for quality control of Bosutinib or its pharmaceutically acceptable salt or its solvates or its hydrates.
According to another aspect, there is provided a highly pure Bosutinib or its pharmaceutically acceptable salt or its solvates or its hydrates thereof substantially free of impurity.
As used herein, “highly pure Bosutinib or its pharmaceutically acceptable salt or its solvates or its hydrates thereof substantially free of impurity” refers to Bosutinib or a pharmaceutically acceptable salt thereof comprising above mentioned impurity or its salt thereof in an amount of less than about 0.15% as measured by HPLC. Specifically, the Bosutinib, as disclosed herein, contains less than about 0.10%, more specifically less than about 0.05%, still more specifically less than about 0.02% of impurity, and most specifically is essentially free of impurity.
One aspect of the present invention is to provide a method of testing the purity of a sample of Bosutinib or its pharmaceutically acceptable salt or its solvates or its hydrates or a pharmaceutical dosage form comprising Bosutinib, which method comprises assaying the said sample for the presence of compound of formula A or formula B or formula C or formula D or formula E using the novel isolated and prepared reference compounds.
Another aspect of the present invention is to provide a method of testing the purity of a sample of Bosutinib or its pharmaceutically acceptable salt or its solvates or its hydrates or a pharmaceutical dosage form comprising Bosutinib, which method further comprises using a sample of compound of formula A or formula B or formula C or formula D or formula E having a purity level of at least 80% as a reference marker.
Brief description of Drawings:
Figure 1: XRPD of Form-Al of Bosutinib.
Figure 2: XRPD of Amorphous Form of Bosutinib.
Detailed description of the invention:
According to one aspect, there is provided a Bosutinib compound, 4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methylpiperazin-1-yl)propoxy]quinoline-3-carbonitrile, or its pharmaceutically acceptable salt or its solvates or its hydrates thereof.
The acid addition salts of Bosutinib can be derived from a therapeutically acceptable acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, propionic acid, oxalic acid, mandelic acid, succinic acid, maleic acid, fumaric acid, methanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, citric acid, and tartaric acid.
According to another aspect, there is provided an impurity of Bosutinib compound of formula A, formula B, formula C, formula D and formula E.
The Bosutinib impurities are isolated, synthesized and characterized. In one embodiment, the impurities can be used as a reference standard for determination of the purity of Bosutinib or a pharmaceutically acceptable salt or its solvates or its hydrates thereof. The impurities are detected and resolved from Bosutinib by HPLC. The structure of the compound of formula A, formula B, formula C, formula D and formula E are deduced by using different analytical techniques.
According to another aspect, there is provided isolated Bosutinib impurities.
The present inventors have surprisingly found that the impurities are formed as during the synthesis of Bosutinib due to chemical instability of the target substance and due to possible side reactions. The impurities are isolated, synthesized and characterized. The method preparation and characterization data is describe below in the examples.
Extensive experimentation was carried out by the present inventors to reduce the level of the impurities in Bosutinib. As a result, it has been found that the where the impurities formed in the preparation of the Bosutinib can be reduced or completely removed.
In addition to the presence of impurities, the solid state physical properties of an active pharmaceutical ingredient (API), can be very important in formulating a drug substance, and can have profound effects on the ease and reproducibility of formulation.
According to another aspect, there is provided a highly pure Bosutinib or a pharmaceutically acceptable salt or its solvates or its hydrates thereof substantially free of impurities.
This invention provides a high yielding, high purity, operational simple process for the production of Bosutinib, comprising:
a. combination of the compound of formula-VII with cyano acetic acid by using reagents which provides good leaving group to obtain the cyano acetamide compound of formula-VIII

wherein suitable good leaving groups including R-N2+ dinitrogen, R-OR'2+ dialkyl ether, R-OSO2RF perfluoroalkylsulfonates, R-OTs tosylates, R-OMs mesylates, R-I iodide, R-Br bromide, R-OH2+, R-OHR water, alcohols, R-Cl chloride
b. reacting compound of formula III with formula IX in presence of alkali halides and base with suitable solvent provides compound of formula-IV

wherein suitable alkali halides including Sodium chloride, Sodium iodide, Sodium bromide, Sodium floride, potassium chloride, potassium iodide, potassium bromide, potassium floride, lithium chloride, lithium iodide, lithium bromide.
Exemplary inorganic bases include, but are not limited to, hydroxides, carbonates, alkoxides and bicarbonates of alkali or alkaline earth metals. Specific inorganic bases are sodium hydroxide, calcium hydroxide, magnesium hydroxide, potassium hydroxide, lithium hydroxide, sodium carbonate, potassium carbonate, lithium carbonate, sodium tert-butoxide, sodium isopropoxide and potassium tert-butoxide, and more specifically sodium hydroxide, potassium hydroxide, sodium carbonate and potassium carbonate. Even more specifically potassium carbonate.
Specific organic bases are organic amine bases of formula NR1R2R3 wherein R1, R2 and R3 are each independently hydrogen, C1-6 straight or branched chain alkyl, aryl alkyl, C3-I0 single or fused ring or independently R1 , R2 and R3 combine with each other to form C3-7 membered cycloalkyl ring or heterocyclic system containing one or more heteroatom
c. reduction of compound of formula-IV in presence of Pd catalyst and suitable alcoholic solvent gives the compound of formula-V

Exemplary alcohol solvents include, but are not limited to, C1 to C6 straight or branched chain alcohol solvents such as methanol, ethanol, propanol, butanol, amyl alcohol, hexanol, and mixtures thereof. Specific alcohol solvents are methanol, ethanol, isopropyl alcohol, and mixtures thereof, and most specific alcohol solvent is ethyl alcohol
d. condensing compound of formula-V with compound of formula-VIII in presence of trialkyl ortho formate, suitable solvent provides compound of formula-VI

Suitable solvents include but not limiting to alcohols, ethyl acetate. Specific alcohol solvents are methanol, ethanol, isopropyl alcohol, and mixtures thereof, and most specific alcohol solvent is isopropyl alcohol
e. cyclodehydration of compound of formula-VI in presence of phosphorus oxychloride, and suitable solvent provides compound of formula-I

f. crystallizing or purifying the crude Bosutinib in presence of suitable solvents to provide suitable crystalline Form.

In an embodiment the suitable solvent is step e, is acetonitrile and is carried out in presence of TBAB.
The detailed process for the preparation of Bosutinib as describes below.

The process can produce Bosutinib crystalline particles in substantially pure form.
The Bosutinib must be analyzed for purity, typically by UPLC, HPLC or GC analysis, to determine if it is suitable for continued processing or ultimately for use in a pharmaceutical product. The five novel impurities of Bosutinib are isolated, synthesized and characterized, which are mentioned as below.
(i) 4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-propoxyquinoline-3-carbonitrile, impurity-A, (Formula-A) which has the following structure:

(ii) 1,4-bis[3,3'-[4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxy-7-propoxyquinoline-3-carbonitrile]]piperazine, impurity-B (Formula-B) which has the following structure

(iii) 1,3-Bis[7,7'-[4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxyquinoline-3-carbonitrile]]propanediol,Impurity-C (Formula-C) which has the following structure:

(iv) (2Z)-2-cyano-N-(2,4-dichloro-5-methoxyphenyl)-3-(4-methylpiperazin-1-yl)prop-2-enamide, Impurity-D (Formula-D) which has the following structure:

(v) N,N'-bis(4-methoxy-3-(3-(4-methylpiperazin-1-yl)propoxy)phenyl)imidoformamide, Impurity-E (Formula-E) which has the following structure:

This invention also provides an operational simple process for the production of compound of formula A, formula B, formula C, formula D, formula E with high yield and high purity.
The following examples are given for the purpose of illustrating the present disclosure and should not be considered as limitation on the scope or spirit of the disclosure.
EXAMPLES
Example: 1 Process for preparation of N-(2,4-dichloro-5-hydroxyphenyl)acetamide

m-Aminophenol (100 g), Acetic acid (500 ml) are charged at 25-30 oC and stirred for 20-30 mins. Acetic anhydride (98.14 g) to the solution at 25-30 oC for 1-2 hrs and stirred for 1 hr. in Acetic acid at 25-30oC, stirred for 2 hours. Monitoring the reaction and added with additional acetic anhydride if necessary. Raising the temperature to 37 ± 2oC, Sulphuryl chloride (253.55g) was added to the reaction mass slowly for about 8 hrs and stirring the reaction mixture for 1 hr, monitoring the reaction and adding Sulphuryl chloride if necessary. Water was added at 8 ± 2oC and stirred for 1 hour, filtered, washed and dried.
Example: 2 Process for preparation of 2,4-dichloro-5-methoxyaniline

Acetone (300ml),N-(2,4-dichloro-5-hydroxyphenyl)acetamide (100g) and K2CO3 powder (81.53g) are charged in to a reaction vessel at 25 ± 5oC, Dimethyl sulphate (62.99g) at 25 ± 5oC was added for 1-2 hrs and stirred for 1 hr. Monitoring the reaction and added with K2CO3 powder (3.7g) and Dimethyl sulphate (2.86g) if necessary, and follow the above steps till the completion of reaction. Water is added, stirred, filtered, washed and suck dried. The obtained wet cake was added with water and conc HCl and stirred, temperature is raised to 103 ± 2oC (Reflux) under stirring and stirred for 4 hrs. Cooled, filtered, the wet cake and water were charged to reaction vessel, adjusting the pH by ammonia solution and stirred, filtered and dried to obtain title compound.
Example: 3 Process for preparation of 2-(3-chloropropoxy)-1-methoxy-4-nitrobenzene

Reaction mass of 2- methoxy-5-nitrophenol (100 g), Potassium carbonate (81.6g) and DMF (250 ml) was stirred for 1 hour at 25-30 oC; to this reaction mass, 1-Bromo-3-chloropropane (186.2 g) was added and the temperature is raised to 36-38 oC and maintained for 12 hrs. Add cyclohexane (500 ml) and maintain the solution for 1-2 hrs at 32±3oC under stirring, followed by addition of process water (1000ml) under stirring conditions (1-2 hrs), then cooling to 27±3oC and stirring for 2 hrs at 27±3oC. Filtered and washed with cyclohexane and water, followed by drying.
Example: 4 Process for preparation of 1-[3-(2-methoxy-5-nitrophenoxy)propyl]-4-methylpiperazine hydrochloride

A reaction vessel was charged with 2-(3-chloropropoxy)-1-methoxy-4-nitrobenzene (100g), acetonitrile (700 ml) and Potassium carbonate (56.24 g), Potassium iodide (13.5 g) as catalyst and NMP (48.92 g) at 25±5oC and stirred. Temperature was raised to 82±3oC (reflux) and maintained for 15 hrs. Cooling the reaction mass to 60oC, distilling out acetonitrile under vacuum, then charged with ethyl acetate and maintained at 52±3oC for 1 hour and cooling to 12±3oC and was stir for 1 hour, solid was filtered and washed with ethyl acetate. Add IPA.HCl at 76±2oC during 30-60 minutes and maintaining it for 27±3oC for 2 hrs, The wet cake was treated with Sodium hydroxide in Methanol, distilled, the residue taken in Ethyl acetate, cooled and filtered the salt. The filtrate was treated with IPA.HCl, cooled, stirred, filtered and dried the 1-[3-(2-methoxy-5-nitrophenoxy)propyl]-4-methylpiperazine hydrochloride (OR optionally 1-[3-(2-methoxy-5-nitrophenoxy)propyl]-4-methylpiperazine dihydrochloride).
Example: 5 Process for preparation of 2-cyano-N-(2,4-dichloro-5-methoxyphenyl)acetamide

Cyanoacetic acid (62.0 g) and MDC were charged at around 27oC, the reaction mass was cooled to 7 ±3oC and then charges with p-TsCl (134.1 g) and stirred for 1 hr. 2,4-dichloro-5-methoxyaniline (100 g) solution in MDC and TEA were added in a separate vessel and added to above cyanoacetic acid solution at 7 ± 3oC in more than 8 hrs under stirring and stirred for 1 hr. Added with water, stirred, filtered, washed with MDC, and dil ammonia solution, and dried to obtain title compound.
Example: 6 Process for preparation of 2-Cyano-N- (2, 4-dichloro-5-methoxyphenyl)-3- {4-methoxy-3- [3- (4-methylpiperazin-1yl)-propoxy]-phenylamino}-acrylamide

The 1-[3-(2-methoxy-5-nitrophenoxy)propyl]-4-methylpiperazine hydrochloride (100 g) was treated with Sodium hydroxide in Methanol, distilled, the residue taken in Ethyl acetate, cooled and filtered. The filtrate was taken for reduction over 10% Pd/C (3.59) under 2-4Kg/cm3 H2 pressure at 34-40oC, after completion of reaction, catalyst filtered and distilled out completely. The residue was reacted with Triethyl orthoformate and 2-cyano-N-(2,4-dichloro-5-methoxyphenyl)acetamide in IPA at 83±2oC, reflux. After completion of reaction, cooled, filtered and dried.
Example: 7 Process for preparation of Bosutinib.

2-Cyano-N- (2, 4-dichloro-5-methoxyphenyl)-3- {4-methoxy-3- [3- (4-methylpiperazin-1yl)-propoxy]-phenylamino}-acrylamide (100g), Toluene (700ml), TBAB (30g) and POCl3 (83.9g) were added in to the reaction flask at RT. Temperature was raised to 107±3oC and maintained for 18 hrs, monitoring the reaction and adding additional POCl3 if needed. Reaction mass was cooled, water was added at 32±3oC and stirred for about 1 hr, separate layers, product obtained was in aq. Layer. Charged with activated carbon, filtered, the clear filterate was added 30 % Sodium hydroxide solution; acetonitrile was added at 42±3oC and stirred at this temperature for 2 hrs. Cooled, filtered, washed and dried. Purified and recrystallized from ethanol-methanol mixture to obtain Bosutinib Form-Al.
Example: 8 Process for preparation of polymorphic Form-V Bosutinib.

The obtained product from above example or the crude Bosutinib or Bosutinib Form-Al (100 g) and Denatured Ethanol (400ml) added to reaction flask and stirred at 27±3oC, temperature is raised to 78±2oC (Reflux) and maintained till the solid dissolves. Activated carbon (5 g) was charged to the reaction mass at 78±2oC (Reflux) and maintained at 78±2oC (Reflux) for 30-45 minutes. Filtered and washed with ethanol, Cool the reaction mass to 62±3oC. Methanol (200ml) was added to the reaction mass for more than 30 minutes at 62±3oC and maintained at 62±3oC for 30 minutes, cooled, stirred for 2 hrs at 27±3oC; Solid obtained was filtered and washed with ethanol-methanol mixture. Cyclohexane (1000 ml) was added in to that solid at RT, temperature was raised to 83±2oC (Reflux) under stirring and maintained for 2 hrs. Cyclohexane is distilled out and reaction mass was cooled, filtered, washed with cyclohexane and dried under nitrogen to obtain Bosutinib Form-V.
Example: 9 Process for preparation of amorphous Form of Bosutinib.
Alternatively, the pure Bosutinib was dissolved in MDC, distilled under vacuum, cooled, charged Cyclohexane, filtered and dried to get Amorphous form.
Example:10 Process for preparation of 4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-propoxyquinoline-3-carbonitrile (Impurity-A)

5-Nitro-2-methoxyphenol was reacted n-Propyl bromide in presence of Potassium carbonate in DMF at 50-55oC for 7 hours. Charged Cyclohexane, cooled, added water, stirred, filtered and dried at 55-60oC.
Yield: 90%, Purity by HPLC: 99.92%, Mass: 212.2 (M + H).
1-Methoxy-4-nitro-2-propoxybenzene was reduced over 10% Pd/C in Ethylacetate at 35oC and 4-5 kg Hydrogen gas pressure in autoclave for 6 hours. After completion of reaction, pressure released, cooled, filtered the catalyst and distilled out solvent.
Yield: 87.68%, Purity by HPLC: 96.55%, Mass: 182.2 (M + H).
4-Methoxy-3-propoxyaniline was reacted with 2-Cyano-N-(2, 4-dichloro-5-methoxyphenyl) acetamide in presence of Triethylorthoformate in IPA at 82oC for 10 hours. After completion of reaction, cooled, filtered and dried at 55-60oC.
Yield: 75.5%, Purity by HPLC: 99.78%, Mass: 450.3 (M - H).
The uncyclised desNMP impurity was heated with Phosphorous oxychloride in Acetonitrile at 90oC for 25 hours. After completion of reaction distilled, cooled, added water followed by neutralization with diluted sodium hydroxide solution, filtered and dried at 55-60oC.
Yield: 100%, Purity by HPLC: 98.17%, Mass: 432.2 (M - H).
Example:11 Process for preparation of 1,4-bis[3,3'-[4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxy-7-propoxyquinoline-3-carbonitrile]]piperazine (Impurity-B)

2-(3-Chloropropoxy)-1-methoxy-4-nitrobenzene was reacted with Piperazine in Acetonitrile in presence of Potassium carbonate and Potassium iodide at 82oC for 41 hours. After completion of reaction, distilled, added water, filtered and dried at 55-60oC.
Yield: 41.87%, Purity by HPLC: 84.79%, Mass: 505.5 (M + H).
Bis 1,4-[3-(2-methoxy-5-aminophenoxy) propyl] piperazine was reduced over 10% Pd/C in Ethylacetate at 28oC and 4-5 kg Hydrogen gas pressure in Parr apparatus for 15 hours. After completion of reaction, pressure released, cooled, filtered the catalyst and distilled out solvent.
Alternatively, 7-(3-chloropropoxy)-4- [(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy quinolone -3-carbonitrile was reacted with Piperazine in Acetonitrile in presence of Potassium carbonate and Potassium iodide at 82oC for 58 hours. After completion of reaction, distilled, added water, filtered and dried at 55-60oC. The crude was purified in Ethylacetate – Cyclohexane – Acetone.
Yield: 42.28%, Purity by HPLC: 94.06%, Mass: 947.5 (M + H).
Example:12 Process for preparation of 1,3-Bis[7,7'-[4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxyquinoline-3-carbonitrile]]propanediol (Impurity-C)

5-Nitro-2-methoxyphenol was reacted 1, 3-Dibromo propane in presence of Potassium carbonate in DMF at 50-55oC for 7 hours. Charged Cyclohexane, cooled, added water, stirred, filtered and dried at 50-55oC.
Yield: 40.58%, Purity by HPLC: 97.36%, Mass: 379.2 (M + H).
1, 1’-[Propane-1,3-diylbis (oxy)]bis (2-methoxy-5-nitrobenzene) was reduced over 10% Pd/C in Ethylacetate at 35oC and 4-5 kg Hydrogen gas pressure in autoclave for 12 hours. After completion of reaction, pressure released, cooled, filtered the catalyst and distilled out solvent.
Yield: 89.13%, Purity by HPLC: 92.7%, Mass: 319.3 (M + H).
3, 3’-[Propane-1,3-diylbis(oxy)] bis (4-methoxyaniline) was reacted with 2-Cyano-N-(2, 4-dichloro-5-methoxyphenyl) acetamide in presence of Triethylorthoformate in IPA at 82oC for 37 hours. After completion of reaction, cooled, filtered and dried at 50-55oC.
Yield: 74%.
The uncyclised propoxy dimer impurity was heated with Phosphorous oxychloride in Acetonitrile at 90oC for 17 hours. After completion of reaction distilled, cooled, added water followed by neutralization with diluted sodium hydroxide solution, filtered and dried at 55oC. Purified in IPA.
Yield: 71.78%, Purity by HPLC: 88.0%, Mass: 821.3 (M + H).
Example:13 Process for preparation of (2Z)-2-cyano-N-(2,4-dichloro-5-methoxyphenyl)-3-(4-methylpiperazin-1-yl)prop-2-enamide(Impurity-D)

N-Methyl piperazine was reacted with 2-Cyano-N-(2, 4-dichloro-5-methoxyphenyl) acetamide in presence of Triethylorthoformate in IPA at 82oC for 14 hours. After completion of reaction, cooled, filtered and dried at 50-55oC.
Yield: 57%, Purity by HPLC: 99.34%, Mass: 369.3 (M+), PMR: (CDCl3): 2.361 (s, 3H), 2.560-2.545 (m, 4H), 3.575-3.544 (t, 2H), 3.917 (s, 3H), 4.054-4.023 (t, 2H), 7.373 (s, 1H), 7.854 (s, 1H), 8.226 (s, 1H), 8.274 (s, 1H).
Example: 14 Process for preparation of N,N'-bis(4-methoxy-3-(3-(4-methylpiperazin-1-yl)propoxy)phenyl)imidoformamide (Impurity-E)

4-Methoxy-3-[3-(4-methylpiperazin-1-yl)propoxy]aniline was reacted with 2-Cyano-N-(2, 4-dichloro-5-methoxyphenyl) acetamide in presence of Triethylorthoformate in IPA at 82oC for 14 hours. After completion of reaction, distilled, charged Cyclohexane, cooled, filtered and dried at 50-55oC.
Yield: 32.01%, Purity by HPLC: 64.61%, Mass: 569.6 (M + H).
,CLAIMS:1. A process of preparation of Bosutinib compound of formula –I comprising:

a) reacting the compound of formula-VII with cyano acetic acid by using p-TsCl to obtain the cyano acetamide compound of formula-VIII;

b) reacting Compound of Formula III with formula IX in presence of Potassium Iodide and K2CO3, in a suitable solvent provides compound of formula-IV;

c) reducing the compound of formula-IV in presence of Pd catalyst and suitable solvent gives the compound of formula-V;

d) condensing compound of formula-V with compound of formula-VIII in presence of trialkyl ortho formate, in a suitable solvent provides compound of formula-VI;

e) cyclodehydrating the compound of formula-VI in presence of phosphorus oxychloride, TBAB and a suitable solvent provides compound of formula-I;

f) Crystallizing or purifying the crude Bosutinib in presence of suitable solvent system to provide suitable crystalline Form.
2. The process according to claim 2, wherein in step f, the solvent is selected ethanol, methanol, cyclohexane and mixture thereof.
3. A process for preparation of crystalline Form-V, comprising treatment of Bosutinib with ethanol-methanol mixture followed by cyclohexane.
4. An Isolated compound having chemical name
a) 4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-propoxyquinoline-3-carbonitrile and structure of Formula-A

b) 1,4-bis[3,3'-[4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxy-7-propoxyquinoline-3-carbonitrile]]piperazine and structure of Formula-B

c) 1,3-Bis[7,7'-[4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxyquinoline-3-carbonitrile]]propanediol and structure of Formula-C

d) (2Z)-2-cyano-N-(2,4-dichloro-5-methoxyphenyl)-3-(4-methylpiperazin-1-yl)prop-2-enamide and structure of Formula-D

e) N,N'-bis(4-methoxy-3-(3-(4-methylpiperazin-1-yl) propoxy) phenyl) imidoformamide and Formula-E

5. The isolated compound according to claim 4 is used as a reference marker in testing the purity of a sample of Bosutinib.