`
`‘Pharmazie 12
`
`D 20485
`
`An International Journal
`Volume 56
`of Pharmaceutical Sciences_December 2001
`
`
`
`In this issue
`
`Pharmaceutical applications of supercritical carbon dioxide
`(Review)
`-
`
`e Histamine H3-receptor antagonists
`
`¢ Optimization of capillary electrophoretic analysis
`e Capillary electrophoretic analysis of sesquiterpenes
`from Valeriana
`
`Fluid-bed melt granulation
`Phosphatidylcholine peroxidation in multilamellar liposomes
`Evaluation of an intranasal formulation of midazolam
`Osmotic pumptablets of naproxen sodium
`Penetration rates of different magnesium salts
`e Inhibition of neutrophil elastase by phenolic compounds
`
`are
`
`Editors
`
`P. Pflegel (Greifswald)
`Editor-in-chief
`
`T. Dingermann (Frankfurt am Main)
`Editor-in-chief
`
`H. Derendorf (Gainesville)
`Editor North America
`
`A. Helmstiidter (Eschborn)
`
`Editorial advisory board
`
`A. Arancibia (Santiago, Chile)
`M.Brewster (Beerse, Belgium)
`W. Charman (Victoria, Australia)
`M. Dittgen (Jena, Germany)
`G. Franz (Regensburg, Germany)
`H. Gebler (Hannover, Germany)
`G. Heinisch (Innsbruck, Austria)
`H.-D. Holtje (Dtisseldorf, Germany)
`O. Hoshino (Tokyo, Japan)
`J. Jurenitsch (Wien, Austria)
`G. Lee (Erlangen, Germany)
`V. Linck Bassani (Porto Alegre, Brazil)
`T. Loftsson (Reykjavik, Iceland)
`H. Morck (Eschborn, Germany)
`K. Mohr (Bonn, Germany)
`R. H. Miiller (Berlin, Germany)
`E. Mutschler (Frankfurt, Germany)
`P. Nuhn (Halle, Germany)
`H.-H. Otto (Greifswald, Germany)
`C. Reppas (Athens, Greece)
`A. Sakr (Cincinnati, USA)
`P. C. Schmidt (Tiibingen, Germany)
`W. Schunack (Berlin, Germany)
`W. Siegmund (Greifswald, Germany)
`J. P. Surmann (Berlin, Germany)
`R. Vistelle (Reims, France)
`
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`Pharmazie 56 (2001) 12
`AQUESTIVE EXHIBIT 1127 Page 0002
`AQUESTIVE EXHIBIT 1127 Page 0002
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`

`

` 2
`
`|
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`>ES
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`aCc&
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`c2=g&=ooe3
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`oO
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`ORIGINAL ARTICLES
`
`Department of Ophthalmology’, National University Hospital, Faculty of Pharmacy?,
`National University Hospital, University of Iceland, Reykjavik, Iceland
`
`and Department of Anaesthesiology’,
`
`Intranasal administration of midazolam in a cyclodextrin based
`formulation: bioavailability and clinical evaluation in humans
`
`H. GupmunpspoTtTiA”', J. F. SIGURJONSDOTTIR?, M. Masson2, ©. FuALLDAL3, E. STEFANSSON’ and T. LoFTSSON?
`
`Intranasal administration of midazolam has been of particular interest because of the rapid and reliable onset of action,
`predictable effects, and avoidance of injections. The available intravenous formulation (Dormicum® IV solution from
`Hoffmann-La Roche) is however less than optimal for intranasal administration due to low midazolam concentration and
`acidity of the formulation (pH 3.0—3.3). In this study midazolam was formulated in aqueous sulfobutylether-B-cyclodex-
`trin buffer solution. The nasal spray -was tested in 12 healthy volunteers and compared to intravenous midazolam in an
`open crossovertrial. Clinical sedation effects, irritation, and serum drug levels were monitored. The absolute bioavailabil-
`ity of midazolam in the nasal formulation was determined to be 64 + 19 % (mean + standard deviation). The peak serum
`
`concentration from nasal application, 42 + 11 ng ml~!, was reached within 10—15 min following administration and clin-
`ical sedative effects were observed within 5 to 10 min and lasted for about 40 min. Intravenous administration gave
`clinical sedative effects within 3 to 4 min, which lasted for about 35 minutes. Mild to moderate, transient irritation of
`nasal and pharyngeal mucosa was reported. The nasal formulation approaches the intravenous form in speed of absorp-
`tion, serum concentration and clinical sedation effect. No serious side effects were observed.
`
`ity of midazolam in the new midazolam nasal spray and
`to determine the clinical sedative effects in healthy volun-
`teers.
`
`2. Investigations and results
`
`Mean midazolam serum concentration curves following
`both modes of administration are shown in the Fig.
`All values shown are the mean values + the standard
`deviation of
`the mean. The maximum midazolam
`
`plasma
`concentration was 42.1+411.2ngml~!
`(range
`29-72 ng ml~!), 15.5+7.9min (range 5—30 min)
`fol-
`lowing intranasal administration. Area under the curve di-
`vided by the dose (in mg) of midazolam administered
`(AUC/dose) was calculated to be 1981 + 487 ng m7!
`min mg!
`following intravenous
`administration
`and
`1209 + 279 ng ml“! min mg™!
`following intranasal ad-
`ministration. The approximate 95% confidence interval
`was from 1705 to 2257 ng ml“! min mg! dose and from
`1052 to 1367ng ml~' min mg™! dose for the AUC fol-
`lowing intravenous and intranasal midazolam, respectively.
`
`
`
`1. Introduction
`
`Due to their well-defined anxiolytic properties, benzodi-
`azepines are among the most popular sedatives for use
`before surgery and anaesthesia. Midazolam, a short-acting,
`water-soluble benzodiazepine, has been demonstrated to
`be a safe and effective preanesthetic anxiolytic agent [1]
`and its clinical efficacy is well documented after intrave-
`nous [2, 3], intramuscular [4], intranasal [5—7], oral [8, 9]
`and rectal [10] administration.
`.
`The available parental drug formulation (Dormicum® IV
`solution from Hoffmann-La Roche) is however less than
`optimal for intranasal use due to the low concentration of
`midazolam as well as the acidic pH (pH 3.3) of the for-
`mulation [11]. Nasal
`irritation [12], discomfort within
`the throat area as well as bitter taste are common com-
`plains following the intranasal administration. The large
`volume needed and accordingly extended administration
`time add to the unpleasant side effects from the avail-
`able parenteral formulation when used for intranasal appli-
`cation.
`By formulating midazolam in a solution containing sulfo-
`butylether-B-cyclodextrin and hydroxypropyl methylcellu-
`lose, the solubility of midazolam (and therefore the mida-
`zolam concentration in the dosage form) was increased
`dramatically [13]. This enhanced solubility could be ob-
`tained at a higher pH (pH 4.3) than previously reported
`for the parenteral drug formulation used as intranasal mid-
`azolam dosage forms. Cyclodextrins and their derivatives
`have been extensively studied for their use as pharmaceu-
`tical excipients. Constructed of a-1,4 linked glucose units,
`they form cone shaped cylinders with hydrophilic exterior
`surface and a hydrophobic inner cavity. This allows cyclo-
`dextrins to form inclusion complexes with a lipophilic
`moiety of the midazolam molecule, increasing the aqueous
`solubility of midazolam dramatically without affecting the
`drug’s pharmacological properties [13—15]. The addition
`of small amounts of water-soluble polymers, such as hy-
`droxypropyl methylcellulose,
`to the solution further in-
`creases the solubility of midazolam in the aqueous cyclo-
`dextrin solution [16, 17]. The main objectives of the
`present study were to determine the absolute bioavailabil-
`
`Pharmazie 56 (2001) 12
`
`150
`
`200
`
`250
`
`300
`
`350
`
`400
`
`Time (min)
`
`Fig: Serum concentration-time profiles following administration of mida-
`zolam intranasal (©) (0.06 mg/kg) and intravenous (Ml) (2 mg). Each
`point represents the mean + standard deviation
`
`
`AQUESTIVE EXHIBIT 1127 Page 0003 |
`AQUESTIVE EXHIBIT 1127 Page 0003
`
`963
`
`

`

`
`ORIGINAL ARTICLES
`
`the intranasal group. Similarly, there werestatistically sig-
`nificant differences (p < 0.05) in sedation from baseline
`from 3 min through 40 min post administration in the in-
`travenous group. Comparison of difference of sedative ef-
`fects between the intranasal and intravenous groups using
`Mann-Whitneytest, revealed statistically significant differ-
`ences (p < 0.05) from 3 to 4 min and from 40 to 50 min
`post administration (Table 1).
`Noclinically significant symptoms of a midazolam ovr
`dose like respiratory depression or cardiovascular depres-
`sion were seen in any of the volunteers. No volunteer
`complained of nausea or vomiting but approximately one
`half of the subjects reported mild to moderate discomfort
`and/or pain within either the nasal passage or throat area
`following intranasal midazolam-cyclodextrin administra-
`tion. Comparison of distribution of nose and throat dis-
`comfort scores during and after study-drug administration
`using Wilcoxson’s test, revealed (p < 0.05) significant dif-
`ferences the first 4 min after administration in the nasal
`passage and thefirst 15 min in the throat area respectively
`(Table 2). No clinically relevant changes in arterial oxygen
`saturation, heart rate and blood pressure and following
`both intravenous and intranasal midazolam administrations
`were demonstrated. In both groups, all subjects had recov-
`
`AQUESTIVE EXHIBIT 1127 Page 0004
`
`IN-Values
`
`* Comparison ofdistribution of sedation scores between control (5 min before adm.) and post-administration using Wilcoxon’s test (*P < 0.05)
`! Comparisonofdistribution of sedation scores between groups IV vs. IN using Mann-Whitneytest (!P < 0.05, !!P < 0.01)
`
`This gave a mean intranasal midazolam bioavailability of
`64 +19%. Mean terminal half-life (t}) following intrave-
`nous midazolam administration was 107.8 + 30.9 min and
`128.8 + 54.0 min following intranasal midazolam admini-
`stration. The approximate 95% confidence interval was
`90 min to 125 min and 98 min to 159 min for the ty. va-
`lues following intravenous and intranasal midazolam ad-
`ministration,
`respectively.
`In one half of the group of
`twelve participants the initial sedative effect was seen
`within 5—10 min, and for one additional participant within
`15 min, following intranasal drug administration. Maximal
`sedative effects were attained from 15—20 min and these
`continued until 55—60 min after administration. Five of
`the above mentioned participants showed initial sedative
`effect within 2—3 min following intravenous drug adminis-
`tration and approximate maximal sedative effects were at-
`tained from 3—4 min until 35—40 min after administration.
`The other members of the group, namely five participants,
`experienced minor or no sedative effects following either
`intranasal or intravenous drug administration. Comparison
`of distribution of sedative scores between control and post
`administration midazolam using Wilcoxson’stest, revealed
`statistically significant differences (p < 0.05) from base-
`line from 8 min through 50 min post administration for
`
`.
`
`Table 2: Distribution of nose and throat discomfort scores after intranasal midazolam administration (n = 12)a
`Discom.
`Contr.
`Admin.
`Time in minutes after administration
`
`score
`
`-5
`
`0
`
`1
`
`3
`
`4
`
`5
`
`8
`
`10
`
`15
`
`20
`
`25
`
`30
`
`a N
`
`43
`
`OSE-Values
`
`1
`1
`1
`2
`3
`3
`3
`2
`3
`2
`3
`2
`2
`3
`1
`
`
`
`
`
`
`
`12 5 6 7 7 9 9 12 12 12 120 12ee
`%#
`*
`*
`*
`
`
`
`
`
`
`
`
`
`
`
`
`3
`4
`
`3
`4
`
`5
`
`3
`2
`
`4 3
`
`THROAT-Values
`
`2
`6
`9
`7
`2
`1
`2
`2
`3
`7
`0
`12
`7
`7
`5
`5
`1
`1
`3
`4
`12
`12,
`12
`
`*
`#
`*
`*
`*
`*
`*
`
`* *
`
`Comparison ofdistribution of discomfort scores between control (5 min before adm.) and post-administration using Wilcoxon’s test (*P < 0.05)
`
`964
`
`Pharmazie 56 (2001) 12
`AQUESTIVE EXHIBIT 1127 Page 0004
`
` TV-Values
`
`Table 1: Distribution of sedative scores after intravenous and intranasal midazolam administration (n = 12)
`Time in minutes after administration
`Sedation
`Contr.
`Admin.
`score
`10
`15
`20
`25
`30
`
`40!
`
`451!
`
`50!
`
`35
`
`
`
`

`

`
`
`_—_-e—RwoToeoAlane
`ORIGINAL ARTICLES
`
`ered from sedation and were fully awake and alert within
`150—180 min of the initial drug administration.
`
`3. Discussion
`
`
`
`The nose is a unique route of drug delivery into the body.
`The nasal cavity is highly accessible to sprays and drops,
`there is a large surface area (approximately 160 cm?) [18]
`through which absorption can occur. Intranasal midazolam
`is absorbed through the highly vascularized nasal mucosa
`[19, 20] directly into the systemic circulation, thus bypassing
`first pass hepatic metabolism. This results in a faster peak
`plasma concentration and a higher bioavailability than can
`be achieved after oral or rectal administrations [21, 22].
`The intranasal route has been extensively described and
`the successful use of intranasal midazolam, although still
`unlicensed via this route, has been reported in children
`undergoing dental procedures [7], anaesthesia induction
`[5, 22—25], and minor surgery [9, 26]. In adult patients,
`intranasal midazolam has been used for the management
`of claustrophobia during magnetic resonance imaging
`[27], for endoscopic procedures [28] for dental surgery [6,
`29] and this administration looks promising for short-term
`management of seizures [30, 31].
`This study demonstrates that it is possible to achieve ef-
`fective intranasal delivery of midazolam using a drug de-
`livery system based on cyclodextrin complexation with a
`bioavailability of the midazolam-cyclodextrin-nasal spray
`of 64 + 19%. In a previous study in six healthy volunteers
`the bioavailability of the nasal spray was determined to be
`73 &+7% [13]. There are some indications that the abso-
`lute bioavailability of midazolam from the nasal cavity is
`dose dependent,
`ie.
`that
`the bioavailability increases
`somewhat with increasing dose but this study was the first
`clinical trial for this nasal-spray formulation and therefore
`the dose was kept at a minimum (0.06 mg/kg). Compara-
`tive pharmacokinetic studies in adults demonstrate that
`Sulfobutylether-B-cyclodextrin was kindly donated by CyDex Inc. (USA),
`bioavailability after oral administration is from 44% to
`hydroxypropyl methylcellulose 4000 was obtained from Mecobenzon
`(Denmark) and midazolam from Sifa (Ireland). All other chemicals used
`68% [32, 33] (depending on dose given) andasingle re-
`were of pharmaceutical grade (European Pharmacopeia, 3rd Edition,
`port of pharmacokinetic data following buccal administra-
`1997). The midazolam nasal spray was formulated in a sulfobutylether-B-
`tion of 5 mg midazolam in adults shows a bioavailability
`cyclodextrin-hydroxypropyl methylcellulose aqueous solution. Midazolam
`of 74.5% [34]. Another single report of pharmacokinetic
`base (1.7 g) was added to 100 ml of an aqueoussolution containing 14 g
`sulfobutylether-8-cyclodextrin, 0.1 g hydroxypropyl methylcellulose, 0,02 g
`data following intranasal administration of midazolam
`benzalkonium chloride, 0.1 g ethylenediaminetetraacetic acid and 0.73 g
`(0.15 mg/kg body weight) in adults demonstrates that un-
`phosphoric acid. The pH of the formulation was adjusted to pH 4.20—4.35
`der optimal conditions absorption of midazolam via the
`with sodium hydroxide. The solution was heated in an autoclave at 121 °C
`nasal mucosa can be virtually complete, with a bioavail-
`for 40 min to promote the complexation [13, 16]. The resulting solution
`wasthen filtered through a 0.45 um membraneand aseptically divided into
`ability of 83% [35] but more often a combined nasal/gas-
`amber crimp cap vials. Finally, the vials and their contents weresterilised
`trointestinal absorption occurs, following intranasal mida-
`in an autoclave at 121°C for 20 min. The nasal spray was prepared at the
`zolam administration, due to the large volume used, with
`facilities of Icelandic Pharmaceuticals (Iceland).
`a bioavailability quoted between 50 and 57% [24, 36].
`This new, more concentrated (17 mg/ml) nasal spray for-
`mulation made it possible to obtain clinically sedative ef-
`fect in adult patients using only 200-300 ul (based on
`body weight) compared with 2—3 ml, using the conven-
`tional
`intravenous (5 mg/ml) solution [29, 36, 37]. The
`sedation levels in our study were evaluated under ideal
`circumstances with no adverse stimuli and the clinical ef-
`fect documented based on the individual sedation assessed
`by the participants. Approximately 10—15 min following
`administration of the nasal spray seven out of twelve parti-
`cipants demonstrated satisfactory sedation status where as
`the other participants experienced minor or no sedative
`effects throughout the study, so clearly the individual re-
`sponses to midazolam plasma concentration differ, accord-
`ing to our study. The same subgroup of participants had a
`clinical sedative effect from both intranasal and intrave-
`nous administration and the other subgroup found no se-
`
`dative effect from every formulation, which indicates the
`individual difference in people’s response to midazolam.
`Additionally a relationship between plasma midazolam
`concentration and clinical effects could not been clearly
`established, following either intranasal or intravenous ad-
`ministration, but some researchers suggest
`that sedation
`may be associated with plasma midazolam concentrations
`greater than 30—100 ng/ml [38, 39].
`The administration time for this new nasal-spray-formula-
`tion was less than 1 min. The majority of the participants
`reported mild to moderate irritation within the nasal pas-
`sage and/or throat area following nasal-spray administra-
`tion. The irritation is most likely due to the bitterness of
`midazolam. Based on a preliminary double blind/rando-
`mised study six volunteers could not differentiate between
`, Saline and the vehicle administered intranasally by a Pfeif-
`fer, “unit dose” device. Evaluation of subjective irritation
`is always difficult but discomfort level in our study was
`scored mild to moderate that is not as severe as previously
`reported [9, 12, 40]. No other side effects were observed
`in the nose and local symptoms such as sneezing and
`coughing were not observed in this study.
`In conclusion, the pharmacokinetic data presented in our
`study demonstrate that the midazolam-cyclodextrin nasal
`spray formulation approaches the intravenous form in
`speed of absorption and serum concentration. By formulat-
`ing midazolam in a solution containing sulfobutylether-p-
`cyclodextrin and hydroxypropyl methylcellulose, the solu-
`bility is increased dramatically at a higher pH (pH 4.3)
`and this unique property, as well as the ease of administra-
`tion, offers
`significant advantages over currently used
`treatment modalities for sedation.
`
`4. Experimental
`4.1 Materials and methods
`
`4.2. Study protocol
`
`The protocol was an open crossover trial, approved by the local ethics
`committee of the National University Hospital and the State Committee on
`Pharmaceuticals in Iceland. Before enrolment, all volunteers gave written
`informed consent. Twelve healthy volunteers were chosen among 18 Cau-
`casian students who applied for participation, on the basis of normalliver
`and kidney function, as reflected in normal creatinine and bilirubin blood
`values and no history or signs of a cold within the two weeksprior to the
`first experimental day. They were prohibited from using any drugs with
`known metabolic interactions with midazolam such as tranquilizers or alco-
`hol 2 days prior to and during the study. The mean age of the volunteers
`was 25.8 years (range 19-37 years) and the mean weight was 73.0 kg
`(range 55-92 kg). Five female and seven male volunteers were included in
`this study. Volunteers reported to the study unit at 10:00 AM each study
`day after an 8-hour fasting period. They continued to fast until 2h after
`administration of the study drug. Electrocardiography and blood oxygena-
`tion monitors were affixed and an intravenous cannulainserted into a fore-
`arm vein for collection of blood samples. Baseline blood samples were
`obtained 5 min prior to administration of the study medication, according
`to protocol. The participants received either intravenous orintranasal appli-
`cation in the first part of the study and the other application was carried
`
`965
`AQUESTIVE EXHIBIT 1127 Page 000
`AQUESTIVE EXHIBIT 1127 Page 0005
`
`5U
`
`T
`
`Pharmazie 56 (2001) 12
`
`

`

`III,
`
`ORIGINAL ARTICLES
`
`Table 3: Rating scale for evaluation of discomfort in mucous
`membranes in nose and throat
`
`range and the 95% confidence interval were also presented. The SPSS
`statistical program wasused to analyze theresults.
`
`Score
`
`Discomfort level
`
`0
`]
`2
`3
`4
`
`Nodifference between nasal spray and saline
`Minor discomfort
`Some discomfort
`Major discomfort
`Burning pain
`
`Table 4: Rating scale for evaluation of sedative effect
`
`Score
`
`Behaviour and signs
`
`Classification
`
`Acknowledgements: The authors would like to thank Professor Magnus
`Johannsson for his invaluable assistance. This study was supported by The
`Icelandic Research Fund for Graduate Students and the University of Ice-
`land Research Fund.
`
`References
`
`1 Kupietzky, A.; Houpt, M.: Pediatr. Dent. 15, 237 (1993)
`2 Barker. I; Butchart, D.; Gibson, J.; Lawson, J.; Mackenzie, N.: Br. J.
`Anaesth. 58, 371 (1986)
`3 Clark, M.; Silverstone, L.; Coke, J.: Oral Surg. Oral Med. Oral Pathol.
`3, 127 (1987)
`4 Rita, L.; Seleny, F; Mazurek, A.: Anesthesiology 63, 528 (1985)
`5 Karl, H.; Keifer, A.; Rosenberger. J.; Larach, M.; Ruffle, J.: Anesthe-
`siolgy 76, 209 (1992)
`6 Fukuta, O.; Braham, R.; Yanase, H.; Atsumi, N.; Kurosu, K.: J. Clin.
`Pediatr. Dent. 17, 231 (1993)
`7 Fuks, A.; Kaufman, E.; Ram, D.; Hovav, S.; Shapira, J.: Pediatr. Dent.
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`Normalreaction.
`(6) — Normal
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`
`Irritable with body movement. +8)—Excited
`18 Gizurarson, S.: Acta Pharm. Nord. 2, 105 (1990)
`20)
`—
`t
`
`Highly irritable with considerable (7,8)—Excite
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`6
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`885 (1993)
`
`1
`
`2
`
`3
`
`4
`
`5
`
`Sleeping. No responseto patting
`on the shoulder.
`
`Sleeping. No reponseto calling by
`nametwoorthree times.
`Respondsto patting on the shoulder.
`
`Eyesclosed, dull reaction.
`Responds when addressed.
`
`Eyes open and closed by turns,
`dull reaction. Responds to verbal
`timulus.
`stuns
`Eyes open, dull reaction.
`Respondsto verbal stimulus.
`
`(1,2) - Asleep
`
`(3) — Drowsy
`
`(4,5) — Sedated
`
`out 7 days later. For intravenous administration the volunteers received
`2mg Dormicum® (IV solution from Hoffmann-La Roche) via an injection
`into a peripheral vein in the contralateral arm. To guarantee a complete
`washout of midazolam between investigations a period of at least 7 days
`was maintained between the two forms of administration. For intranasal
`administration the volunteers received 0.06 mg/kg (193-323 pl, based on
`body weight) dose of midazolam-cyclodextrin via a nasal spray, with half
`of the study druginstilled rapidly into one nostril, and the remainder into
`the other nostril over 1 min. The spray bottle device, “Unit dose” closed
`system, from Pfeiffer, delivered a fine aerosol and was held at an angle of
`approximately 45° to the vertical with the nozzle held at an angle of ap-
`proximately 5° to the sagittal plane, pointing slightly towards the nasal
`septum and introduced as far as possible into the nostrils without causing
`pain. There was somespillage of the midazolam dose in one subject. Since
`the amount spilled could not be quantified, this subject was omitted from
`the calculations for peak concentration (Cp max) and the time to reach peak
`level (tmax)- After administration of study medication (0h) blood samples
`(5 ml) were collected from the intravenous catheter at 5, 10, 15, 20, 30,
`60, 120, 180, 240, and 360 min. Samples were centrifuged for 10 min, and
`serum harvested and frozen at —20°C until analyzed for midazolam con-
`centration. At each sampling point the volunteers were questianed regard-
`ing nasal mucosa irritation (Table 3) and degree of sedation (Table 4).
`Vital signs,
`including heart rate, blood pressure, oxygen saturation and
`respiratory rate were monitored continuously and registered at 5 min inter-
`vals during 2h after drug administration. Serum samples were assayed for
`midazolam by reverse-phase HPLC at
`the Medicinsk Laboratorium A/S
`(Copenhagen, Denmark). Standard methods were used to calculate the
`pharmacokinetic parameters. The serum level from intravenous and intrana-
`sal application was compared for each participant. The maximum midazo-
`lam serum concentration (Cp max) and the time to reach it (tmax) were ob-
`tained from the serum concentration profiles. The area-under-curve (AUC)
`was calculated using the linear trapezoidal method, the absolute bioavail-
`ability (F) was calculated as the percent of AUCintanasal over AUCintravenous
`ratio within the O~360 min time period.
`
`4.3. Statistical analysis
`Results were analyzed using the Student’s t-test (paired-samples t-test and
`independent-samplet-test) and the non-parametric tests Wilcoxon’s t-test (2
`related samples test) and-Whitney U-test (2 independent samplestest). The
`level of significance was set at p< 0.05. The mean, standard deviation,
`
`Received June 8, 2001
`Accepted July 4, 2001
`
`Thorsteinn Loftsson
`Faculty of Pharmacy
`University of Iceland
`P.O. Box 7210
`127 Reykjavik
`Iceland
`thorstlo@hi.is
`
`Pharmazie 56 (2001) 12
`AQUESTIVE EXHIBIT 1127 Page 0006
`AQUESTIVE EXHIBIT 1127 Page 0006
`
`