Enhancing the physicochemical properties of solid-state materials through crystal engineering enables optimization of these materials without covalent modification. Cocrystals have become a reliable means to generate novel crystalline forms with multiple components and they exhibit different physicochemical properties compared to the individual components. This dissertation exemplifies methodologies to generate cocrystals of active pharmaceutical ingredients (API's) based upon knowledge of supramolecular interactions (supramolecular synthons), while focusing on enhanced delivery through in vitro and in vivo processes with both salts and cocrystals respectively. The utility of mechanochemistry involving small amounts of an appropriate solvent, or solvent drop grinding (SDG), has been shown to reliably reproduce cocrystals with the anti-convulsant carbamazepine that were originally obtained by solution crystallization. This technique has been confirmed as a reliable screening method using solvents in which both components exhibit some solubility. The benefits of this technique lie in the time and cost efficiency associated with it as well as its inherently small environmental impact making it a "Green" method. SDG was also used as an efficient way to discover cocrystals of the anti-inflammatory meloxicam with carboxylic acids after analysis of existing reports and the analysis of structural data from the Cambridge Structural Database (CSD) to guide the choice of coformer. It has been shown that SDG can be used to screen for cocrystalline forms that are also obtainable by solution crystallization which is important in later stage development and manufacturing including but not limited to large scale up processes. Single crystals suitable for single crystal X-ray diffraction were obtained with meloxicam and two of the coformers, fumaric and succinic acid. Some of the meloxicam cocrystals exhibited enhanced pharmacokinetic (PK) profiles in rats exemplifying significantly higher serum concentrations after only fifteen minutes and consistently higher exposure over the time studied while others maintained lower exposure. This reveals that cocrystals can fine tune the PK profile of meloxicam in order to reduce or enhance exposure. Two different sulfonate salts, 4-hydroxybenzenesulfonate (p-phenolsulfonate) and 4-chlorobenzenesulfonate, of the anti-spastic agent (R,S) baclofen were developed by strategically interrupting the intramolecularly stabilized zwitterionic structure of baclofen. This zwitterionic structure results in low solubility associated with physiological pH required for intrathecal administration. Structural data for both salts in the form of single crystal X-ray diffraction data was successfully obtained. Solubility based on baclofen was assessed and shown to increase in pure water and at pH's 1 and 7. Only the 4-chlorobenzenesulonate salt maintained an increased solubility over two days at pH 7 making it a viable candidate for further study in terms of intrathecal administration. During crystallization experiments with (R,S) baclofen two polymorphic forms of the baclofen lactam were generated, Forms II and III. Both forms are conformational polymorphs confirmed by single crystal X-ray diffraction and Form II has a Z' of 4 with an unusual arrangement of enantiomers.

Ischemic injury occurs when the flow of blood is reduced or blocked to an area of the body and can cause significant tissue damage by generation of reactive oxygen species (ROS), activation of apoptotic pathways and through induction of the inflammatory response. Restoration of blood flow and reperfusion of the blocked site, while essential, can generate a second injury that itself needs to be controlled. Together the two injuries are termed ischemia/reperfusion (I/R) injury. This type of injury is frequently encountered in medicine and is a major medical problem. Therapeutic strategies to combat I/R injury include the introduction of compounds that can scavenge ROS or can induce metabolic pathways with the effect of inhibiting apoptosis. Caffeic Acid Phenethyl Ester (CAPE), a polyphenolic compound found in propolis, has been shown to protect a variety of cells types against ROS in vitro and has also been shown to induce a variety of genes including hemeoxygenase 1 (HMOX-1) , an enzyme that has been implicated in a cytoprotective pathway. Despite showing significant cytoprotection of cells against oxidant stress in vitro, CAPE is readily hydrolyzed in plasma and is also quickly removed from circulation. This result may explain the limited cytoprotective effects of CAPE in vivo. We have synthesized a series of CAPE amide derivatives, including Caffeic Acid Phenethyl Amide (CAPA), with the aim of improving CAPE’s stability properties while maintaining the cytoprotective effects of the parent compound. We found that CAPA, in addition to 2 other amide derivatives, were able to protect human umbilical vein endothelial cells (HUVEC) against ROS to a similar degree as CAPE. In addition, we have observed significant improvement in plasma stability of CAPA over CAPE at multiple temperatures. The elimination half-life of CAPA from the systemic circulation was also seen to be significantly improved over CAPE following intravenous administration to male Sprague-Dawley rats. The longer residence time of CAPA over CAPE in circulation may potentially result in greater cytoprotection in vivo. / text

published_or_final_version / Statistics and Actuarial Science / Doctoral / Doctor of Philosophy

Thesis (MTech. degree in Pharmaceutical Sciences)--Tshwane University of Technology, 2010. / Investigates the absorption enchancing properties of N-trimethyl chitosan chloride (TMC) and monocaprin (MC), individually and in combination, on the permeability of insulin across the Caco-2 intestinal epothelial cell line.

Η θεραπεία με λεβοθυροξίνη (LT4) έχει στενό θεραπευτικό παράθυρο. Απόκλιση δοσολογίας μπορεί να οδηγήσει σε υπό-θεραπεία ή σε τοξικότητα. Οι δόσεις υποκατάστασης ή καταστολής λεβοθυροξίνης εξαρτώνται τόσο από την απορρόφηση της λεβοθυροξίνης, όσο και από την άλιπη μάζα σώματος. Όσον αφορά τις θέσεις απορρόφησής της από το γαστρεντερικό σωλήνα, οι γνώσεις μας προέρχονται από περιορισμένες μελέτες χορήγησης ραδιοσημασμένης θυροξίνης. Σκοπός της παρούσης εργασίας ήταν να καθορίσει τις φαρμακοκινητικές παραμέτρους της LT4 σε άτομα με νοσογόνο παχυσαρκία, να συγκρίνει αυτές με τις αντίστοιχες νορμοβαρών ατόμων και να μελετήσει τις θέσεις απορρόφησης της λεβοθυροξίνης στο γαστρεντερικό σωλήνα εκμεταλλευόμενοι το πλεονέκτημα των βαριατρικών επεμβάσεων στους ασθενείς με κλινικά σοβαρή παχυσαρκία, στις οποίες αφαιρείται άλλοτε άλλο τμήμα του γαστρεντερικού σωλήνα. Μέθοδος. Μελετήσαμε 62 ευθυρεοειδικά άτομα με αρνητικά αντισώματα κατά της θυρεοειδικής υπεροξειδάσης, 38 ασθενείς με νοσογόνο παχυσαρκία (ομάδα SOS) και 24 υγιείς εθελοντές (ομάδα αναφοράς). Οι 32 ασθενείς από την ομάδα SOS, υπεβλήθησαν σε επιμήκη γαστρεκτομή (10 άτομα), σε γαστρική παράκαμψη κατά Roux-en-Y (7 άτομα) και σε χολοπαγκρεατική εκτροπή με δημιουργία μακρών ελίκων (15 άτομα). Σε όλα τα άτομα χορηγήθηκε από το στόμα διάλυμα 600mcg νατριούχου λεβοθυροξίνης, μετά από ολονύκτια νηστεία, και μετρήθηκαν οι τιμές των Τ4 και T3 του πλάσματος στους χρόνους 0, 0.5, 1, 1.5, 2, 2.5, 3 και 4 ώρες μετά τη χορήγηση του διαλύματος. Η ίδια διαδικασία ακολουθήθηκε και 35 ημέρες μετά το χειρουργείο. Αποτελέσματα. Οι φαρμακοκινητικές παραμέτροι Επιφάνεια Κάτω από την Καμπύλη και Μέγιστη Συγκέντρωση της Τ4 ήταν χαμηλότερες μετά τη χορήγηση του διαλύματος LT4 (p<0,01 και p<0,05), ενώ ο Χρόνος Μέγιστης Συγκέντρωσης της Τ4 μεγαλύτερος στην ομάδα SOS σε σύγκριση με την ομάδα των νορμοβαρών (p<0,01). Μετεγχειρητικά στην ομάδα που υπεβλήθη σε επιμήκη γαστρεκτομή, η ΕΚΚ της Τ4 ήταν μεγαλύτερη (p<0,01), ενώ η ΜΣ και ο ΧΜΣ ήταν παρόμοιοι πριν το χειρουργείο και 35 ημέρες μετά το χειρουργείο. Στην ομάδα που υπεβλήθη σε γαστρική παράκαμψη κατά Roux-en-Y, μετεγχειρητικά η ΕΚΚ, η ΜΣ και ο ΧΜΣ της Τ4 ήταν παρόμοια. Στην ομάδα που υπεβλήθη σε χολοπαγκρεατική εκτροπή και δημιουργία μακρών ελίκων, η ΕΚΚ και η ΜΣ της Τ4 ήταν μεγαλύτερες μετεγχειρητικά (p<0,001), ενώ ο ΧΜΣ ήταν παρόμοιος. Συμπεράσματα. Άτομα με νοσογόνο παχυσαρκία πιθανώς να χρειάζονται μεγαλύτερες δόσεις καταστολής ή υποκατάστασης με LT4 σε σχέση με νορμοβαρή άτομα εξαιτίας, εκτός των άλλων παραγόντων, επηρεασμένων φαρμακοκινητικών παραμέτρων της λεβοθυροξίνης.. Οι φαρμακοκινητικές παράμετροι της απορρόφησης της λεβοθυροξίνης είναι βελτιωμένες μετά από βαριατρική επέμβαση. Η λεβοθυροξίνη απορροφάται κύρια από το απώτερο τμήμα της νήστιδας και από τον ειλεό, ενώ το στομάχι και το δωδεκαδάκτυλο δεν εμπλέκονται άμεσα στην απορρόφηση της λεβοθυροξίνης. / Levothyroxine (LT4) has a narrow therapeutic window so that slightly changes in the treatment dose could have significant clinical consequences. Suppressive or replacement doses of LT4 are affected by the rate and extend of the active ingredient absorbed, as well as by the lean body mass. Concerning the absorption sites of levothyroxine, the data are limited and our knowledge based on studies of radio isotopic thyroxine in intestinal tract. Aim of this study was to determine the pharmacokinetic parameter of LT4 in severely obese individuals, compare them with similar data in lean control subjects, as well as to study the consequences of bariatric surgery on the pharmacokinetic parameters of LT4 and to identify the parts of the gastrointestinal tract implicated in its absorption. Method. We studied 62 euthyroid subjects, who had negative tests for anti-thyroid peroxidase antibodies. Thirty eight of these were severely obese but otherwise healthy (SOS-group). Twenty four were healthy controls subjects (control group). Thirty two subjects of the SOS group, underwent sleeve gastrectomy (SG; n=10); Roux-en-Y gastric bypass (RYGBP;n=7) or biliopancreatic diversion with gastric bypass and long limbs (BPD-LL;n=15). Subjects received 600mcg oral solution of sodium LT4 after an overnight fast. Serum T4 and T3 were measured 0; 0.5; 1; 1.5; 2; 2.5; 3 and 4 hours after LT4 administration. The same procedure was repeated 35 days after surgery. Results. The Area Under the Curve (AUC) and the peak plasma Concentration (Cmax) of T4 after LT4 administration were lower; whereas the Time to reach Cmax (Tmax) from the baseline was higher in the SOS than in the control group. Following surgery; in the SG group; the mean AUC was higher (p<0.01); whereas Cmax and Tmax were similar to pre surgery. In the RYGBP group; mean AUC; Cmax und Tmax were similar. In the BPD-LL group; mean AUC and Cmax were higher 35 days after surgery than before (p<0.001); whereas Tmax was similar. Conclusions. Severely obese individuals may need higher LT4 suppressive or replacement doses than normal weight individuals due, among other factors, impaired LT4 pharmacokinetic parameters. The pharmacokinetic parameters of LT4 absorption are improved following bariatric procedures that result in malabsorption.T4 is absorbed primarily from lower part of the jejunum and from the ileum, whereas stomach and duodenum are not involved directly in the absorption of LT4.

Inter-individual variability in drug response is a significant clinical concern and may lead to the development of adverse drug reactions, which are currently a top-ten cause of death in the United States. Recently, the manifestation of disease, which may alter normal physiological function, has gained increased attention for its role as a contributing factor in the development of inter-individual responses to drugs. One such disease, known as nonalcoholic fatty liver disease (NAFLD), is the most common chronic liver disease in Western society and represents a spectrum of clinical morbidities that range from the usually benign simple fatty liver to the more advanced nonalcoholic steatohepatitis (NASH). Prior investigations have identified liver-specific alterations in xenobiotic transporter and metabolizing enzymes in NASH, which lead to the functional disruption of drug disposition. To identify a useful model(s) that is representative of hepatic transporter expression profiles in humans with NASH, gene and protein expression profiles of liver membrane transporters were assayed across several commonly used experimental rodent models of the disease. NASH models that were representative of the human condition developed global, adaptive changes in transporter regulation in the liver, which was not present in models that failed to recapitulate human profiles. Specifically, decreased expression of hepatic uptake transporters was coupled with an induction of efflux transporters, which may serve as a hepatoprotective response by limiting hepatic exposure to potentially harmful substances during times of tissue stress. To link a possible molecular mechanism for these hepatic adaptations in NASH, the role of the oxidative stress-activated transcription factor, Nrf2, was investigated. A functional Nrf2 regulatory element was identified within the eighth intron of the human ABCC3 transporter gene, implicating Nrf2 activation in NASH as a contributor to the coordinated induction of hepatic efflux transporters in the disease. To further clarify the effects of NASH on renal membrane transporter regulation, a thorough analysis of gene and protein expression was conducted with the validated rodent models used previously. Following the manifestation of disease, a global induction of renal efflux was observed, suggesting a compensatory, coordinated response of membrane transporters in the kidney upon disease induction. The functional consequences of liver and kidney xenobiotic transporter dysregulation was shown to disrupt the disposition of the environmental toxicant, arsenic. Specifically, NASH results in increased excretion of arsenic into urine as well as altered hepatic and renal exposure. These findings are associated with hepatic and renal transporter dysregulation and demonstrate for the first time that NASH alters the disposition of environmental toxicants. In summary, these studies contribute novel findings that identify liver and kidney-specific adaptations in disease that may contribute to global alterations in xenobiotic disposition thereby increasing the likelihood of developing adverse drug reactions in patients with NASH.

Studies in this dissertation were conducted to explore the roles of organic cation transporters (OCTs) in the disposition of N-butylpyridinium Chloride (NBuPy-Cl) and structurally related ILs. Following a single i.v. dose to rats, the blood concentration of NBuPy-Cl and 1-butyl-1-methylpyrrolidinium chloride (BmPy-Cl) decreased in a biphasic manner with a clearance of 3.3 and 7 ml/min, respectively. More than 84% of dosed compounds were excreted in the urine. Depending on the vehicle, the dermal absorption of BmPy-Cl and NBuPy-Cl (5 mg/kg, 125 μg/cm²) was 10-35% at 96 h. Following a single oral (50 mg/kg) administration to rats, the maximum blood concentrations of both ILs were reached in less than 90 min in rats. Most of the orally dosed NBuPy-Cl (62-68 %) was excreted in the urine in 72 h. However, more of the dosed BmPy-Cl was eliminated in the feces Its oral bioavailability was only 47%. The elimination differences between BmPy-Cl and NBuPy-Cl were not altered by the size (0.5, 5, or 50 mg/kg) or frequency (1 or 5 administrations) of oral doses. In all urine and blood samples, only parent compounds were detected. Co-administration of NBuPy-Cl and inulin intravenously to rats revealed that the clearance of NBuPy-Cl exceeded the rat glomerular filtration rate, suggesting a renal secretion processing. The in vitro transport studies demonstrated that NBuPy-Cl, BmPy-Cl and 1-butyl-3-methylimidazolium chloride are substrates (Kt, 9~277 μM), as well as inhibitors (IC₅₀: 0.2~7.5 μM), of rOCT1/2 and hOCT2. Their inhibitory effects increased dramatically with increasing the alkyl chain length. The IC₅₀ values were 0.1, 3.8, 14 and 671 μM (hexyl-, butyl-, ethyl-pyridinium and pyridinium chloride) for rOCT2 mediated metformin transport. Similar structurally related inhibitory kinetics were observed for rOCT1 and hOCT2. In vivo co-administration of NBuPy-Cl prolonged the plasma half-life and reduced renal clearance of the diabetic drug, metformin. In summary, BmPy-Cl and NBuPy-Cl are partially absorbed from gastrointestinal tract. The present in blood is eliminated rapidly in the urine as parent, by renal filtration and OCT-mediated secretion. ILs also compete with other substrates of OCTs and have the potential to alter their pharmacokinetic profiles.

Tulathromycin is a macrolide antibiotic approved for use in cattle and swine respiratory disease. Extra-label use of tulathromycin occurs in bison and deer and significant interspecies differences in pharmacokinetics warrant specific investigation in these species. This study involved investigation of the pharmacokinetics of tulathromycin in bison and white-tailed deer following a single 2.5 mg/kg bw subcutaneous injection (n=10) of Draxxin (Pfizer Inc.) to provide important information regarding tulathromycin dosage regimens in these species. As well, tulathromycin distribution and depletion in deer muscle and lung tissues following a 2.5 mg/kg bw subcutaneous injection of Draxxin was investigated to obtain pilot information regarding withdrawal time of tulathromycin in deer. For the pharmacokinetic studies, serial blood samples were collected at baseline and up to 25 days post-injection. Pharmacokinetic parameters were estimated using non-compartmental methods. For the tissue pilot study, deer (n = 2 to 3) were slaughtered at 0, 1, 2, 6, 7, and 8 weeks post-injection. A quantitative analytical liquid chromatography-mass spectrometry method for measuring tulathromycin was developed and validated in bison and deer serum and deer lung and muscle according to international guidelines. Samples were processed by solid-phase extraction. Reverse-phase chromatography was performed by gradient elution. Positive electrospray ionization was used to detect the double charged ion [M+2H]+2 at m/z 403.9 and monitored in selected ion monitoring mode. Tulathromycin demonstrated early maximal serum concentrations, extensive distribution, and slow elimination characteristics in deer and bison. In bison, mean Cmax (195 ng/mL) was lower compared to cattle (300 to 500 ng/mL) and half-life (214 hours) longer (cattle, 90 to 110 hours). In deer, mean Cmax (359 ng/mL) is comparable to cattle, but half-life (281 hours) was much longer. Tissue distribution and clinical efficacy studies are needed in bison to confirm extensive distribution of tulathromycin into lung and the appropriate dosage regimen. Tulathromycin was extensively distributed to deer lung and muscle, with tissue levels peaking within 7 to 14 days after injection. Drug tissue concentrations were detected 56 days after treatment, longer than the established withdrawal time of 44 days in cattle. This prolonged drug concentration in the tissue is supportive for the administration of tulathromycin as a single injection therapy for treatment of respiratory disease of deer. While more study is needed to establish a recommended withdrawal time, the long serum and tissue drug half-life and extensive interindividual variability in tissue levels suggests a withdrawal period well beyond 56 days may be required in deer.

This study examined the effects of 30 minutes of cycle exercise at 65% V̇O2max on the pharmacokinetics of the S.S.R.I. sertraline. Blood samples were taken over 48 hours from 14 healthy males (23.9±2.5 years, 80.3±12.6 kilograms) following oral ingestion of a single 100 mg dose of sertraline. Participants completed two sertraline trials separated by at least two weeks; one trial while resting and the other trial with exercise as described above. With exercise, the absorption rate constant and volume of sertraline in the central compartment decreased, while the elimination half-life increased. Maximum concentration, time of maximum concentration, and area under the curve were unchanged. Fitness level had little impact on the concentration of sertraline, as compartmental modeling was unchanged when relative V̇O2max was added as a covariate. However, controlling for participant body weight improved the model estimate. These results indicate that acute aerobic exercise has the potential to change the concentration of sertraline in vivo.

No description available.