Department of Pharmaceutical Analysis, Pullareddy Institute of pharmacy, Hydarabad, 502313, India
A novel sensitive, accurate and safe UV -Spectrophotometric method was developed for the estimation of anti -cancer drug (bosutinib).The various analytical parameters were validated for the bulk drug according to ICH Q2 guidelines. The proposed method includes two method, Method-A development and validation where methanol were used for all validation studies .As drug is more likely soluble in water it was scanned in the UV where it has shown maximum absorbance at 268nmThe linearity response was observed in the concentration range of 2-12ug/ml,where r2 =0.0996was the regression value the RSD value for method precision was found to be well within acceptance criteria &LOD,LOQ was established for bosutinib further more stability studies for bosutinib were carried out under acidic ,basic andoxidation as per stability assay methods The results of accuracy and recovery studies were carried out by adding specific drug amount(50%,100%,150%)and shown the recovery in range of (96.6%-98.6%).The proposed method was successfully applied for method development,validation and stability studies of bosutinib.
Bosutinib is an antineoplastic agent used for the treatment of chronic, accelerated, or blast phase Philadelphia chromosome-positive (Ph+) chronic myelogenous leukemia (CML).
It is chemically 4-[(2,4-dichloro-5-methoxyphenyl) amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl) propoxyl-3-quinoline carbonitrile
Bosutinib is a 7-alkoxy-3-quinolinecarbonitrile that functions as a potent, dual SRC and ABL tyrosine kinase inhibitor indicated for chronic myelogenous leukemia (CML), specifically Philadelphia chromosome-positive (Ph+) CML. Philadelphia chromosome is a hallmark of
CML due to the reciprocal translocation t(9; 22) (q34;q11) , resulting in a BCR-ABL fusion protein.The first BCR-ABL inhibitor, imatinib, was introduced over a decade ago as a breakthrough in CML management;however, emerging resistance to imatinib poses challenges in achieving remission.4 Second- generation BCR-ABL inhibitors like bosutinib inhibit most resistance-conferring BCR-ABL mutations except V299L and T315, thus providing more therapeutic options for patients.
DRUG PROFILE3
Bosutinib was first approved by the FDA in 2012 for the treatment of adult chronic, accelerated, or blast-phase Ph+ CML with resistance or intolerance to prior therapy. On September 26, 2023, bosutinib was also approved by the FDA for the treatment of pediatric CML that is newly diagnosed or resistant/intolerant to prior therapy.
Fig 1: Chemical Structure Of Bosutinibmesylate
IUPAC NAME:
4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methyl-1-piperazinyl) propoxy]-3-quinolinecarbonitrile.
MOLECULAR FORMULA: C26H29C12N5O3.
MECHANISM OF ACTION:
Bosutinib is a tyrosine kinase inhibitor. Bosutinib inhibits the BCR-ABL kinase that promotes CML; it is also an inhibitor of Src-family kinases including Src, Lyn, and Hck. Bosutinib inhibited 16 of 18 imatinib-resistant forms of BCR-ABL kinase expressed in murine myeloid cell lines. Bosutinib did not inhibit the T315I and V299L mutant cells
USES:
Bosutinib is used to treat a certain type of chronic myeloid leukemia (CML; a type of cancer of the white blood cells)
MATERIALS AND INSTRUMENTS
Materials used:
Instrument used
Table-1: Instrument
SOLUBILITY STUDIES AND SELECTION OF SOLVENT:
Table-2: Solubility Studies
SELECTION OF SOLVENT:
After Solubility Studies we have selected Methanol as the best solvent, because compared to other solvents Bosutinib was immediately dissolved in methanol
MAXIMUM WAVELENGTH:
? max: The wavelength at which a substance has its strongest photon absorption (highest point along the spectrums Y- max) .
Fig 2: ? Max Of Bosutinib
Table-3: Linearity
METHODOLOGY4-17
Method development and validation for the estimation of bosutinib by using UV- spectrophotometer
Preparation of standard solution:
Standard stock 1:
10.0mg of Bosutinib was accurately weighed and transferred to a 10ml volumetric flask. A few ml of methanol was added to dissolve the drug and the volume was made up to the mark with methanol which gives the concentration of 1000µg/ml.
Standard stock II:
1.0ml from standard stock I was pipetted to a 10ml volumetric flask and the volume was made up to the mark with methanol which gives the concentration of 100µg/ml.
VALIDATION OF THE DEVELOPED METHOD:
The developed UV method was validated for precision, linearity, accuracy, LOD, and LOQ.
A series of linear concentrations (2 - 12µg / ml) were prepared from standard stock II solutions Appropriate dilutions were made from the aliquot into separate 10 ml volumetric flasks and made up to the volume with methanol. Noted down the absorbance for each of the concentrations (Table no: 9) and a plot of the calibration curve was constructed (shown in Fig.3)
INTRADAY:
The intraday precision was carried by scanning the sample at different times within a day. %RSD is found to be 0.501%
Acceptance criteria: The % RSD for the intraday precision is calculated and the calculated value is within the limits, i.e.?2%.
INTERDAY:
The interday precision was carried out by scanning the sample at different days with in a week. %RSD is found to be 0.504%
Acceptance criteria: The %RSD for the Interday precision is calculated and the calculated value is within the limits, i.e.,?2%.
ACCURACY:
Recovery of the spiked standard solution (known concentration) was performed at three different levels (0%, 100%, and 150%) of the sample solution. The % recovery was then calculated from the recorded absorbance values. (Table-7)
LIMIT OF DETECTION:
Least amount of concentration to be detected
Formula: 3.3 x standard deviation/mean
Value found:
1.962µg
LIMIT OF QUANTIFICATION:
Least amount of concentration that is to be quantified
Formula: 10 x standard deviation/mean
Value found:
5.945µg.
DRUG STABILITY STUDIES:
Acid and Base Degradation:
1ml from Standard stock II (100?g/ m l) was pipetted to a 10ml, volumetric flask, and 2ml.of 0.1N HC1/0.IN NaOH was added. After 24hrs neutralize the solution with 2ml of 0.IN NaOH/0.IN HCl the remaining volume were made up to the mark with water (10ug / ml) 2ml each of Appropriate 0.1N HCI and 0.1N NaOH were taken into respective 10ml volumetric flasks and made up to the mark with water. These were used as blank. The absorbance of the acid/base degraded samples was noted applying the calibration developed method.
Oxidation (Hydrogen peroxide):
1ml from Standard stock II (100mg / ml) was pipetted to a 10ml volumetric flask and 2ml of 3% H2O2 was added. After 24hrs the solution is finally made up to the mark with distilled water. The absorbance was measured using the above solution against a blank containing 2ml of 3% H2O2 in a 25ml volumetric flask was kept as blank. The absorbance of the peroxide degraded samples was noted applying the developed method.
RESULT AND DISCUSSION
LINEARITY
Table-4: Calibration Plot Of Bosutinib
Fig 3: Calibration Graph Of Linearity Study
PRECISION
Table-5: Intraday Precision Studies
Table-6: Interday Precision Studies
ACCURACY
Table-7 Recovery Studies
STABILITY STUDIES
Table-8: Degradation Studies
SUMMARY
CONCLUSIONS
ACKNOOWLEDGEMENT;
The authors are very much thankful to Pullareddy institute of Pharmacy ,Hyderabad, Telangana. For providing necessary facilities and continuous support for this entire research work.
CONFLICT OF INTREST
No conflicts of interest.
REFERENCES:
Vardhineedi Shirisha,R. Parasuraman, M. Poojitha, R. Gayathri, P. Mounika, M. Bheemaji, Development And Validation Of Bosutinib By Using UV – Spectrophotometric Method In Bulk Form, Int. J. of Pharm. Sci., 2024, Vol 2, Issue 5, 1544-1550. https://doi.org/10.5281/zenodo.11352665
10.5281/zenodo.11352665
