Upload
others
View
0
Download
0
Embed Size (px)
Citation preview
FORMULATION AND CHARACTERIZATION OF PARENTERAL
DOSAGE FORM CONTAINING AN ANTINEOPLASTIC AGENT BY
LYOPHILIZATION TECHNIQUE
M. Pharm Dissertation Protocol
Submitted to
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA
BENGALURU
By
Mr. RIZWAN AHMED
M.PHARM, PART-I
Under the guidance of
Dr. Dinesh B.S. M.Pharm, Ph.D
Professor
DEPARTMENT OF PHARMACEUTICS
PRIYADARSHINI COLLEGE OF PHARMACYKORATAGERE, TUMKUR
2012-2013
ANNEXURE – II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. Name of the Candidate and Address
RIZWAN AHMED
PRESENT ADDRESS:M.PHARM 1ST YEAR, DEPT OF PHARMACEUTICSPRIYADARSHINI COLLEGE OF PHARMACY,KORATGERE, TUMKUR
PERMANENT ADDRESS: S/O M D HUSSAINNEAR MASJID, MAIN ROAD,POST: KARDAKALTQ:LINGSUGURDIST: RAICHUR
2. Name of the Institute PRIYADARSHINICOLLEGEOF PHARMACY,TUMKUR.
3. Course of the Study and Subject M.PHARMACY IN PHARMACEUTICS
4. Date of Admission to the Course 05 JUL 2012
5. Title of Topic
“FORMULATION AND CHARACTERIZATION OF PARENTERAL DOSAGE FORM CONTAINING AN ANTINEOPLASTIC AGENT BY LYOPHILIZATION TECHNIQUE”
1
6. BRIEF RESUME FOR INTENDED WORK
6.1 Need for the Study
Antineoplastic agents are a group of specialized drugs used primarily to treat cancer.
“Cancer is a disease of uncontrolled cell division, invasion and metastasis. It is generally
considered to be due to the clonal expansion of a single neoplastic cell. However there may
be additional somatic leading to heterogeneous cell population. It is one of the major
causes of death in the developed nations: one in three people will be diagnosed with cancer
during their lifetime.1
The anticancer drugs either kill cancer cells or modify their growth. However, selectivity
of majority of drugs is limited and they are one of the most toxic drugs used in therapy.
Treatment of malignant diseases with drugs is a rather recent development, started after
1940 when nitrogen mustard was used, but progress has been rapid, both in revealing
pathobiology of the diseases and discovery of new drugs. In addition, attempts have been
made to define optimal combinations, treatment strategies and patient support measures.
Cancer chemotherapy is now of established value and a highly specialized field.
In addition to their prominent role in leukemia and lymphoma, drugs are used in
conjunction with surgery, radiotherapy and immunotherapy in the combined modality
approach for many solid tumors, especially metastatic chemotherapy is the initial treatment
of choice in malignant diseases, drugs are used:
To cure or prolonged remission.
To palliation (alleviation of symptoms).
Adjuvant chemotherapy: Drugs are used to mop up any residual malignant
cells (micro metastases) after surgery or radiotherapy.
2
The drugs which are commonly used in treatment of cancer include Methotrexate,
Gemcitabine, Cisplatin, Carboplatin, Azacitidine, Pemetrexed, Bleomycin, Decitabine,
Epirubicine, Mitoxantrone, Ifosfamide, Mitomycin, Dactinomycin and plant alkaloids such
as Vincristine, Vinblastine, Topotecan, Paclitaxel, Docetaxel, Etoposide etc. The main
side effect of chemotherapy includes nausea and vomiting, hair loss, anemia etc. 2
The typical route of administration of anticancer drugs are oral (tablets and capsules) and
parenteral as intramuscular, Subcutaneous and intravenous. Often a pharmaceutical
product may be susceptible to physical and chemical degradation when stored as a ready-
to-use solution. The goal of the formulations scientist is to identify the right formulation
conditions, the right excipients in optimal quantities, and the right dosage form to
maximize stability, biological activity, safety, and marketability of a particular product. In
order to overcome the above listed draw backs, the solution dosage form meant especially
for parenteral use can be formulated as lyophilized products especially with antineoplastic
agents, formulating them into lyophilized product may improve their stability in the dry
state and thus improve shelf-life3.
Lyophilization is a process more commonly known as freeze-drying. The word is derived
from Greek and it means "made solvent-loving". The process of freeze-drying is used often
to stabilize delicate and heat-sensitive materials and compounds such as: proteins,
peptides, biologicals, vaccines, enzymes, microorganisms, monoclonal antibodies,
parenteral, pharmaceuticals, blood fractions, oglionucleotides, blood plasma and
diagnostics blood fractions etc4.
In the formulation of lyophilized products the typically used excipients are the
cryoprotectant and lyoproctant. Usually used cryoprotectant and lyoproctant are trehalose,
3
glucose, mannitol, sucrose and lactose in the concentration of 0.2-5%. These agents are
intended to prevent freezing stress and drying stress on the drug molecule5.
The primary mechanism that allows for freeze-drying is sublimation, whereby ice is
directly converted to water vapor, without passing through the intermediary stage of a
liquid. Rather than through heating, this is done by removal of solvent under pressure so
that the ice boils without melting.
The result is a sample whose structure is largely preserved, which can be stored at room
temperature and pressure. The process consists of three separate, unique, and
interdependent processes; freezing, primary drying (sublimation phase), and secondary
drying (desorption phase)6.
Thus the lyophilization of antineoplastic agents will result in products with following
advantages:
Ease of processing a liquid, which simplifies aseptic handling.
Removal of water without excessive heating of the product.
Enhanced product stability in a dry state.
Rapid and easy dissolution of reconstituted product.
Longer shelf life.
Less cumbersome storage, handling and distribution requirements.
Lower shipping costs because of reduced weight, especially for bulk applications7.
Decitabine (brand name Dacogen®) and pemetrexed (brand name Almita®) are anticancer
drugs belongs to antimetabolites classification available in the market as lyophilized
formulations. Lyophilization process protocols of individual drugs needs to be designed
4
and developed as and when changes are made in formulation ingredients (especially
cryoprotectants). These protocols are to be designed based upon physiochemical and
thermal characterization of individual drugs.
In the present study research work is aimed to develop a suitable injectable dosage form
for the drug candidate by working on designing of suitable lyophilization cycle by varying
the total cycle time, freezing, primary drying and secondary drying time, ramp time,
holding time and keeping all the quantities of all the active pharmaceutical ingredients
constant. Additional attempts will be made to develop scale up studies to reduce cycle time
to meet the industrial requirement. Hence there is a need to develop antineoplastic agents
such as Decitabine, Pemetrexed disodium, Mitomycin, Doxorubicin, Epirubicin,
Oxaliplatin, Melphalan etc. into lyophilized product by designing suitable lyophilization
cycle.
6.2 Review of Literature:
1. Jeffrey Cummings et al., have studied Pharmacokinetics and metabolism of
Mitomycin C (MMC) in NMRI mice bearing MAC 16 colon adenocarcinoma after
direct intratumoural injection of either 500µg free MMC or the same dose
incorporated in albumin microspheres. Microspheres produced a tumor
pharmacokinetic profile of steady state drug levels, avoiding the much higher early
peak (20.5 µg/tumor vs 98.9 µg/tumor) and lower trough of free MMC, and reducing
significantly the levels of drug reaching the systemic circulation (AUC 1.8 µg/ml x
hr for microspheres vs 6.8µg/ml x hr for free drug). 2, 7-Diaminomitosene (2, 7-
DM), a key intermediate in MMC quinone bioreduction, was used as an indicator of
drug metabolic activation in tumor tissue. Peak levels were lo-fold higher (11.2
µg/tumor vs 1.1 µg/tumor) and area under the curve 5-fold higher after free drug.
5
Even taking into account differences in tumor pharmacokinetic profiles of the parent
drug, microspheres actively inhibited 2, 7-DM formation 3-fold. However, the
microspheres generated a completely different pattern of drug metabolism where
four previously uncharacterized mitosane metabolites and elevated levels of cis and
trans 1-hydroxy 2, 7-diaminomitosene were detected. Despite similar parent drug
exposure in tumors, free drug was significantly more active (P < 0.05, Student’s t-
test) against MAC 16. These results suggest that formation of 2, 7-DM’ correlates
more closely with antitumor activity than sustained parent drug levels or appearance
of other key metabolites. Potentially, they provide the first direct evidence for an in
vivo mechanism of action dependent on bioreductive activation and formation of 2,
7-DM8.
2. Klaus Langer et al., have concluded on Freeze drying is a suitable technique to
improve the long-term storage stability of colloidal drug carrier systems such as
nanoparticles. Aim of this study was to systematically evaluate excipients for the
freeze drying and long-term stability of albumin-based nanoparticles. In our study,
nanoparticles made of human serum albumin (HSA) were freeze dried in the
presence of different cryoprotective agents and after reconstitution were evaluated
with regard to their physico-chemical characteristics. Empty, doxorubicin-loaded,
and PEGylated nanoparticles were prepared and were freeze dried in the presence
of different concentrations of sucrose, trehalose, and mannitol, respectively. The
samples were physicochemically characterized with regard to lyophilisate
appearance, particle size, and polydispersity using photon correlation spectroscopy.
For evaluation of long-term stability, the samples were stored at 2–8, 25, and 40ºC
over predetermined time intervals. In the absence of cryoprotectants, particle
growth was observed in all freeze-dried formulations. In the presence of sucrose,
6
mannitol, and trehalose aggregation of HSA nanoparticles during the freeze-drying
procedure was prevented. Although all of the excipients were identified to be
suitable stabilizers for freeze drying of HSA nanoparticles, sucrose and trehalose
were superior to mannitol, especially with regard to the long-term storage stability
results9.
3. Neeraj Kumar et al., have focused on lyophilization process, physicochemical
characterization and long-term storage stability studies of lyophilized doxorubicin-
loaded (PEG) 3–PLA nanopolymersomes. Nanopolymersomes were prepared by
nanoprecipitation method using (PEG) 3–PLA copolymer and lyophilized in the
presence of different lyoprotectants and evaluated for physicochemical properties.
The lyophilized product was studied for long-term stability at 2–8ºC, 25ºC/65%
RH and 40ºC/75% RH over predetermined periods and evaluated for changes in
physicochemical properties. In the absence of lyoprotectants, product was
collapsed with no cake formation. In the presence of inulin, mannitol, gelatin,
polyvinyl alcohol and glycine, nanopolymersome formulations demonstrated an
intact cake occupying same volume as original frozen mass. The residual moisture
content was below 2.5% w/w. The time of reconstitution was instantaneous
following addition of water without any manual shaking. However, inulin found to
be superior to the rest of lyoprotectants for overall lyophilisate physicochemical
properties. Drug loaded nanopolymersome were physically stable and no effect was
observed in terms of physicochemical properties following one year at 2–8ºC. In
conclusion, the results suggest that doxorubicin loaded nanopolymersomes could
be lyophilized using inulin 5% w/v without losing its physicochemical properties
and can be stored at 2–8ºC with a provisional shelf life of more than a year10.
7
4. Valentino J. Stella et al., have evaluated the potential of using (SBE)7m-β-CD and
HP- β -CD as enabling excipients to improve on the current melphalan injectable
formulation. Melphalan is an anti-neoplastic agent formulated for parenteral use as
a sterile, non-pyrogenic, freeze-dried powder. It is marketed by Glaxo-Wellcome as
ALKERAN® for Injection (Alkeran). A major concern with melphalan therapy,
other than its intrinsic cytotoxicity and biocompatibility, arises from its marginal
aqueous solubility and chemical stability; thus, co-solvents are used in the current
two-vial formulation. Because of the two-vial system, the product is also
inconvenient to use. Two approaches to improve melphalan’s formulation utilizing
cyclodextrins, including the use of aqueous (SBE)7m- β -CD or HP- β -CD solutions
as the reconstitution diluents, and: or the use of (SBE)7m- β -CD as a freeze-drying
excipient in a melphalan formulation, are presented. Results showed that, when the
cyclodextrins were used as diluents, the use of organic co-solvents can be
eliminated and the shelf-life of the reconstituted melphalan greatly enhanced.
When the freeze-dried melphalan:(SBE)7m- β -CD formulation was prepared, the
formulation was found to be stable; and a simplified one-vial delivery system was
achieved. In conclusion, the parenterally safe b-cyclodextrins derivatives can
provide promising alternatives and improved formulations for melphalan injectable
and perhaps similar problematic drugs11.
5. Arvind K. Bansal et al., have reviewed on excipients used in various lyophilized
formulations of small molecules. The role of excipients such as bulking agents,
buffering agents, tonicity modifiers, antimicrobial agents, surfactants and co-
solvents has been discussed. Additionally, a decision making process for their
8
incorporation into the formulation matrix has been proposed. A list of ingredients
used in lyophilized formulations marketed in USA has been created based on a
survey of the Physician Desk Reference (PDR) and the Handbook on Injectable
Drugs. Information on the recommended quantities of various excipients has also
been provided, based on the details given in the Inactive Ingredient Guide (IIG)12.
6.3 OBJECTIVES OF THE STUDY :
To achieve the ultimate goal of developing a lyophilized formulation of antineoplastic
agents to improve long term stability, the present work is designed to address the following
objectives:
Preformulation studies on the anticancer drug substance
Formulation of the injectable dosage form
Lyophilization of the injectable dosage form
Development and optimization of lyo-cycle
Evaluation of the lyophilized product as per official and In-house specification
Characterization for drug excipient interaction
Accelerated stability studies
Documentation of resultant data.
9
7.
7.0 MATERIALS AND METHODS
Materials:
Drugs: Any one anticancer drug such as Decitabine, Pemetrexed disodium, Mitomycin,
Doxorubicin, Epirubicin, Oxaliplatin and Melphalan will be chosen at the time of study.
Excipients: Mannitol, lactose, povidone etc.
Method:
Lyophilization technology
7.1. Source of Data
Review of Literature from
a) Journals such as,
1) Drug Development & Industrial Pharmacy
2) Pharmaceutical Development & Technology
3) Journal of Pharmaceuticals Science Technology
4) International Journal of Pharmaceutics
5) Cancer chemotherapy & Pharmacology
6) Pharmaceutical research
7) PDA Journal of Pharmaceutical Science Technology
8) European Journal of Pharmaceutics and Biopharmaceutics
b) Web sites such as
1) drugs.com (www.drugs.com)
2) pubmed (www.ncbi.nlm.nih.gov/pubmed)
3) sciencedirect (www.sciencedirect.com)
10
7.2 Method of Data Collection :
The physiochemical properties of the drug will be collected from drug information center,
various standard books, journals, websites and other sources like research literature data
bases.
The methodology to be followed to achieve the objectives are outlined as follows:
Preformulation studies on drug like nature, color, solubility etc. will be carried out.
Selection of suitable excipients based on compatibility results.
Lyophilization of the injectable dosage form.
Development and optimization of lyo-cycle.
Evaluation of the lyophilized product by different parameters like related
substance, pH, assay, water content, reconstitutional time, reconstitutional stability
etc.
Stability studies will be carried out for developed products as per the ICH
guidelines.
7.3. Does the study require any investigations or investigations to be conducted on
patients or Other humans or animals?
“ NO ”
7.4. Has ethical clearance been obtained from your institution in case of 7.3?
“ Not Applicable”
11
8.
LIST OF REFERENCES:-
1. www.drugs.com/cancer.html (Date of Access 24/11/2012)
2. www.answers.com/topic/antineoplastic-agents (Date of Access 19/11/2012)
3. Frank Kofi Bedu-Addo. 2004. Lyophilization. Pharmaceutical Technology: 10-18.
4. Rey L, May JC. Freeze Drying/Lyophilization of Pharmaceuticals and Biological
Products.2nd Ed. New York. Basel: Marcel Dekker, Inc:2004:1-4
5. Abdelwahed W, Degobert G, Stainmesse S, Fessi H. Freeze Drying of
Nanoparticles Formulation, Process and Storage Considerations. Adv Drug Deliv
Rev.2006; 58:1688-713.
6. Xiaolin (Charlie) Tang and Michael J. Pikal Design of Freeze-Drying Processes
for Pharmaceuticals: Practical Advice. Pharm res. 2004;21(2):191-200.
7. http://www.fda.gov/ICECI/Inspections/InspectionGuides/ucm074909.htm (Date of
Access 10/11/2012)
8. Jeffrey Cummings, Lucy Allan, John F. Smith Encapsulation Of Mitomycin C In
Albumin Microspheres Markedly Alters Pharmacokinetics, Drug Quinone
Reduction In Tumor Tissue And Antitumor Activity Implications For The Drug’s
In Vivo Mechanism Of Action. Biochemical Pharmacology. 1994;47(8):1345-
1956.
9. Marion G. Anhorn, Hanns-Christian Mahler, Klaus Langer. Freeze Drying Of
Human Serum Albumin (HSA) Nanoparticles with Different Excipients. Int. J
Pharm. 2008;363:162-9.
10. Wubeante Yenet Ayen, Neeraj Kumar. A Systematic Study on Lyophilization
Process of Polymersomes for Long-Term Storage Using Doxorubicin-Loaded
(PEG) 3–Pla Nanopolymersomes. European J Pharm Sci. 2012;46:405-14.
11. David Q. Ma, Roger A. Rajewski, Valentino J. Stella. New Injectable Melphalan
12
Formulations Utilizing (SBE)7m-Β-Cd Or Hp-Β-Cd. Int. J Pharm. 1999;189:227-
34.
12. Ankit Baheti, Lokesh Kumar, Arvind K. Bansal. Excipients Used In
Lyophilization Of Small Molecules J. Excipients And Food Chem.2010;1(1):41-
54.
13
9. Signature of the Candidate
(RIZWAN AHMED)
10. Remarks of the Guide: The topic selected for dissertation is satisfactory and feasible
11. 11.1 GuideDr.Dinesh B.S. M.Pharm, Ph.D
Professor & H.O.D,DEPARTMENT OF PHARMACEUTICSPRIYADARSHINI COLLEGE OF PHARMACYKORATAGERE, TUMKUR.
11.2 Signature of Guide:
(Dr.Dinesh B.S.)
11.3 Head of the Department:
Dr. Dinesh B.S. M.Pharm, Ph.D
Professor & H.O.D,DEPARTMENT OF PHARMACEUTICSPRIYADARSHINI COLLEGE OF PHARMACYKORATAGERE, TUMKUR.
11.4 Signature of HOD:
(Dr. Dinesh B.S.)
12. 12.1 Remark of the Principal:
The above mentioned information is found correct and I recommend the same for approval.
12.2 Principal
P.S.MINHAS PRINCIPALPRIYADARSHINI COLLEGE OF PHARMACYTUMKUR
12.Signature of the Principal
(P.S.MINHAS)
14