Therapeutika. fiir Mangelzusta.nde
`Therapeutics for States of Deficiency
`
`Investigation on the Dosage/Efficacy Relationship
`of Iron Dextran in Veal Calves
`
`P. Geisser•, H. Hohl•, M. Baer•, H. Heimb, and W. Fischerc
`
`Summary
`The efficacy of a single dose of 800 mg resp. 1600 mg iron
`in the form of an intramuscularly administrable iron(Ill)(cid:173)
`dextran complex (Anaemex®, CAS 9004-66-4) has been
`tested. On 3 groups of 13 calves each, 0 ml (comparing
`group), 4 ml resp. 8 ml iron dextran 20 % have been ap(cid:173)
`plied. All calves received iron-containing food during the
`test period of 10 weeks. At the beginning of the therapy,
`5 weeks and 8 weeks after application, the parameters:
`weight, hemoglobin, erythrocytes. hematocrit, mean cor(cid:173)
`puscular hemoglobin (MCH), MCH concentration, mean
`corpuscular vol1.1me, plasma protein, fibrinogen, leuko(cid:173)
`c.vtes and serum iron were measured.
`After JO weeks the dead weight has been determined and
`the spleen of some calves tested histologically. The study
`shows that, by the administration of 1600 mg iron as a
`depot injection, a better growth rr;sults with the same qual(cid:173)
`ity of veal. The red-coloring of the veal was not signifi(cid:173)
`cantly different from that of the comparing group. The
`histological findings show especially that the iron depots
`of the spleen were empty in all three groups and thereby
`in this collective no connection exists between the color of
`the veal and the tested dosage of iron dextran 20 %.
`It is considered meaningf1.1I and economic to renounce in
`fut1.1re the iron-containing food and in its place to apply
`intram1.1scular a single dose of 1600-2400 mg iron per
`calf
`The res1.1/ts are compared with a study 011f1.1II term infants,
`which has shown that a intram1.1sc1.1lar single dose of 150
`mg of iron as iron dextran at birth affords a nutritional
`advantage in iron status for up to 15 months. This ad(cid:173)
`vantage is based on the fact that ferrous s1.1/fate interacts
`with food st1.1ffs and therefore the 1.1sed iron fortified milk
`does not f1.1/jill the req1.1irement. From this point of view
`the problems of oral iron absorption and the efficacy of
`intramusc1.1lar iron dextran in the mentioned trial can be
`compared with this st1.1dy.
`
`Zusammenfassung
`Untersuchung zz1r Dosis-Wirkungs-Bezieh1.1ng van Eisen(cid:173)
`Dextran bei Kiilbern
`
`Die Wirkung einer Eimnaldosis van 800 mg b=w. 1600
`mg Eisen in Form eines intramuskuliir verabreichbaren.
`20 % Eisen enthaltenden Eisen(JJI)-Dextran-Priiparates
`(Anaemex®, CAS 9004-66-4) wurde an Kiilbern unter(cid:173)
`sucht. Den Tieren der drei Gruppen van je 13 Kiilbern
`wurden O ml (Vergleichsgruppe), 4 ml b=w. 8 ml van Ei(cid:173)
`sendextran 20 % appliziert. Alie Kiilber erhielten wiihrend
`der Versuchsperiode van 10 Wochen eisenhaltiges Fu/fer.
`Bei Therapiebeginn sowie 5 und 8 Wochen nach Appli(cid:173)
`kation des Eisenpriiparates wurden folgende Parameter
`bestimmt: Gewicht, Hiimoglobin, E1J1thro=yten, Hiima(cid:173)
`tokrit, durchschni1tliches korpusk1.1liires Hiimoglobin
`(MCH), MCH-Konzentration und durchschni1tliches Kor(cid:173)
`puskularvolumen, Plasmaprotein. Fibrinogen, Leuko=.v(cid:173)
`ten und Serumeisen.
`Nach 10 Wochen wurde das Schlachtgewicht registriert
`1.111d van einzelnen Kiilbern die Milz histologisch unter(cid:173)
`sucht. Die Studie zeigt, da/3 bei Applikation van 1600 mg
`Eisen als Depotinjektion ein besseres Wachstum resultiert.
`wobei die Fleischqualitii.t erhalten bleibt. Die Rotfiirbung
`des Fleisches unterschied sich nicht signifikant van der(cid:173)
`jenigen der Vergleichsgruppe. Die histologischen Unter(cid:173)
`such1.1ngen zeigen, da/3 die Eisendepots der Mil= in alien
`drei Gruppen leer waren und da/3 in diesem Kollektiv keine
`Bezieh1.111g zwischen der Rotflirbung des Fleisches und den
`verabreichten Testdosen besteht.
`Es wird als sinnvoll und wirtschaftlich erachtet, =ukii1{ftig
`auf eisenhaltiges Futter zu verzichten und an dessen Stelle
`eine Einmaldosis van 1600 bis 2400 mg Eisen pro Kalb
`intramuskulii.r zu appli=ieren.
`Die Resultate wurden einer Studie mit neugeborenen Kin(cid:173)
`dern gegeniibergestellt, welche zeigt, da/3 eine intramus(cid:173)
`kulii.re Einmaldosis van 150 mg Eisen als Eisendextran
`bei Geburt einen ernii.hrungsmii.j3igen Vorteil beziiglich des
`Eisenstat1.1s bis z1.1 einem Alter van 15 Monaten bringt.
`Dieser Vorteil ist auf die Tatsache zuriick=ufiihren, daj3
`Eisen(JJ)-sulphat mit Lebensmittelbestandteilen Interak(cid:173)
`tionen gibt, zmd da/3 desha/b die venvendete. mit Eisen(ll)(cid:173)
`sulphat angereicherte Milch, den Anforderungen nichl
`vollumflinglich gerecht wird. Von diesem Standpunkt aus
`gesehen konnen die Probleme der oralen Eisenabsorption
`und die Wirksamkeit van intramusku/ii.rem Eisendextran
`der erwii.hnten Studie mit der vorliegenden Studie vergli(cid:173)
`chen werden.
`
`Key words:
`
`Anaemex® · Anti-anaemic drugs · CAS 9004-66-4 · Iron dex/ran, dose-efficacy relationship
`
`Hausmann Laboratories Ltd., Research Departmenta, St. Gallen, Ciba Geigy AG, Section for Animal Healthh. Basel,
`and a veterinary surgeon's practice', Ober/zmkhofen (Switzerland)
`
`32
`
`Arzneim.-Forsch./Drug Res. 41 (I), Nr. I ( 1991)
`Geisser et al. -
`Iron dextran
`
`PGR2020-00009
`Pharmacosmos A/S v. American Regent, Inc.
`Petitioner Ex. 1042 - Page 1
`
`

`

`1. Introduction
`The absorption of iron from food in fattened calves is not
`in every situation so extensively guaranteed that it could
`not lead to anaemia and delay to growth. This lies, on the
`one hand, in the fact that the absorption of iron from the
`gastro-intestinal tract lies in general scarcely above 20 %
`of the dosage, and, on the other hand, together with the
`food usually antibiotics like spiramycine or olaquindox
`are simultaneously given, which can inhibit the absorp(cid:173)
`tion of iron. This is especially the case, when iron salts
`and antibiotics with phenolic groups are combined. In
`addition, iron salts, in contrast to iron complexes, can
`give interactions with food components [4]. These prob(cid:173)
`lems do not exist with iron complexes, which can be ap(cid:173)
`plied intramuscularly or intravenously. These have the
`additional advantage that the absorption and utilization
`is not only at about 20 %, as in case of orally administered
`preparations, but at 90-100 %.
`The dosage-efficacy relationship of iron supply on the
`quality of veal and on the growth of veal calves can be
`better tested or respectively optimised with parenterally
`administered preparations.
`The problems of interaction of iron salts with food stuffs
`are well known in human medicine and lead to the rec(cid:173)
`ommendation that iron(Il)-salts should be taken up to I
`h before meals. But this recommendation is rarely ob(cid:173)
`served in practice. Olivares et al. [7] have shown on full
`term infants that an intramuscular single dose of 150 mg
`of iron as iron dextran (CAS 9004-66-4) at birth affords
`a nutritional advantage in iron status for up to 15
`months, when compared with an unfortified group. The
`group fed an iron fortified milk, containing 15 mg iron
`as ferrous sulfate and 100 mg/! ascorbic acid, however,
`caught up with the iron dextran group by 9 months. From
`this point of view we can compare the problems of oral
`iron absorption and the efficay of iron dextran in our
`study with the study of Olivares.
`
`2. Methods
`2.1. Randomization and test parameters
`In a randomized study the influence of a single dosage of iron
`dextran 20 % on the quality of veal, blood parameters, growth,
`frequency of sickness and side effects was tested on a total of 39
`veal calves. Therefore three groups which were homogenous with
`regard to race, weight and gender, have been formed .
`
`2.2. Dosages of iron dextran 20 %
`and iron requirement
`The iron requirement which is caused by the growth from about
`60 kg to about 170 kg can be calculated as follows at a mean
`haemoglobin level of IO g Hb/ I 00 ml:
`110 kg x 0.07 I/kg x I 00 g Hb/1 x 0.0034 g Feig Hb = 2.618 g Fe
`The contribution of iron-enriched food of the first six weeks is
`only 25 kg x I 00 mg Fe/kg= 2500 mg Fe of which about 500 mg
`Fe are absorbed because the absorption ratio is not higher than
`20 %. In the next four weeks food was given with an iron content
`of 52 mg Fe/kg, which contributes once again to an iron absorp(cid:173)
`tion of about 500 mg Fe.
`The loss of iron amounts to a minimum of 1-2 mg per day,
`which means about I 00 mg Fe for the treating period of 12
`weeks. Therefore a depot dosage of 800 resp. 1600 mg Fe was
`tested.
`
`2.3. Characterisation of iron dextran 20 %
`(batch 958108)1)
`Iron content
`Dextran content
`
`198 mg/ml
`205 mg/ml
`
`1> Anaemex®; manufacturer: Laboratorien Hausmann, St. Gal(cid:173)
`len (Switzerland).
`
`Arzneim.-Forsch./Drug Res. 41 (I), Nr. I ( 1991)
`Geisser et al. -
`Iron dextran
`
`6.0
`pH of the solution
`relative viscosity (H 20 = I) 20. 7
`< 0.16
`KAv-value
`> 10 000 mg Fe/kg b.w.
`LD50 white mice, i.p.
`Iron dextran 20 % is a polynuclear ferric-hydroxide, complexed
`with dextran. The weight average molecular weight Mw, which
`is determined by gel chromatography (KAv-value) in comparison
`to dextran, lies in the region of 180,000 Dalton. In comparison
`to the iron dextran 5 % preparation for human use, described in
`the B.P. (British Pharmacopeia) and the U.S.P. (United States
`Pharmacopeia), the complex stability and the molecular weight
`are higher.
`
`2.4. Administration, scheme of therapy
`and blood sampling
`Iron dextran 20 % was injected into the cervical muscle of the
`veal calves at an age of 3 to 6 weeks.
`Group I: no iron injection.
`Group 2: 4 ml Iron dextran 20 % as a single dose.
`Group 3: 8 ml Iron dextran 20 % as a single dose.
`
`+ + +
`
`6
`
`10 weeks
`
`+
`
`0
`t
`
`4
`
`+ slaughter day
`I blood sampling
`t Iron dextran
`20 % single
`Injection:
`BOO mg Fe (4 ml}
`1600 mg Fe (8 ml}
`
`Scheme 1: Scheme for blood sampling.
`
`Type of food (dosage):
`Provimilk 608 (25 kg) for week 1-6.
`Provimilk 610 (25-50 kg) for week 7-10.
`Composition of food
`Type 608
`Type 610
`Iron content
`I 00 mg/kg
`52 mg/kg
`Spiramycin content
`200 mg/kg
`I 00 mg/kg
`Olaquindox content
`200 mg/kg
`100 mg/kg
`(Manufacturer: Provimi Lacta SA, Cossonay-Gore, Switzer(cid:173)
`land.)
`
`2.5. Histological testing
`The iron staining was done by Berliner blue reaction (on Fe3+)
`and also by Turnbull blue reaction (on Fei+-total iron). Both
`methods gave identical results with test cuttings of an iron(cid:173)
`loaded rat spleen. In the following judgement the tested prepa(cid:173)
`rations were prepared with the Berliner blue reaction.
`
`2.6. Local reactions
`After the injection into the cervical muscle of the corresponding
`amount of iron dextran 20 % the local reaction at the injection
`site is observed during the first three days.
`
`3. Results
`3.1. Parameters of measurement
`The results of the parameters: weight, Hb, Ery, Htk,
`MCH, MCHC, MCV, Plasmaprotein, Fibrinogen, Leu(cid:173)
`kocytes and serum iron are given in Table I and in Fig.
`1-11.
`
`3.2. Weight
`The increase in weight of the three groups is in the same
`manner, whereby numerically and statistically there is no
`difference between the groups 1 and 2. The increase in
`weight of the group 3 with 8 ml iron dextran 20 % as a
`single dosage is on an average of6.5 kg above the increase
`of group 1. This increase is not significant according to
`t-test. According to the sequential analysis for pair dif(cid:173)
`ference the difference in weight on the level 2a = 0.1 is
`significant.
`
`33
`
`PGR2020-00009
`Pharmacosmos A/S v. American Regent, Inc.
`Petitioner Ex. 1042 - Page 2
`
`

`

`m
`
`,;;
`C,
`.c
`"'
`·;;
`
`,00 ...
`,.,
`- ...
`,,.
`~ ... ..
`..
`
`0
`
`10
`
`s
`8
`weeks after treatment
`Fig. 1: Development of weight during the whole test period of IO weeks.
`The mean values and the standard errors of the means (SEM) correspond
`to Table I. • Control, .6. 4 ml iron dextran 20 %, • 8 ml iron dextran 20 %.
`These details apply also to Fig. 2-11.)
`
`••
`
`~
`~
`~ ,.
`.2
`8'
`E .,
`.c
`
`0
`
`5
`weeks after treatment
`Fig. 3: Development of hemoglobin during the first 8 weeks of the test
`period.
`
`e
`
`..
`
`,.
`
`..
`..
`..
`
`E
`'
`~ .,
`C
`]
`e Q.
`E .,
`0 a.
`
`0
`
`0
`
`8
`
`5
`weeks after treatment
`Fig. 2: Development of plasma protein during the first 8 weeks of the test
`period.
`
`..
`..
`..
`..
`
`g
`'c i E
`
`.,
`.c
`
`~,_.-- ----
`~ f ~
`
`0
`
`e
`
`5
`weeks after treatment
`Fig. 4: Development of hematocrit during the first 8 weeks of the test pe(cid:173)
`riod.
`
`Table 1: Result of measurements of the 3 groups of calves at 0, 5, 8 and 10 weeks after application. Dosage: 0 ml, 4 ml resp. 8 ml of iron dextran 20 %
`for groups A, Band C. The mean values, the standard errors of the means (SEM) and the number of calves measured are given.
`
`0
`
`5
`
`8
`
`10
`
`Average
`
`SEM
`
`178.85
`180.92
`183.08
`
`19.72
`20.87
`25.23
`
`n
`
`13
`13
`13
`
`Weight (kg)
`
`Plasma protein (g/1)
`
`Hemoglobin (g/dl)
`
`Hematocrit (%)
`
`Iron (µ1/1)
`
`Erythrocytes (I 06/µl)
`
`Leukocytes ( I 03/µl)
`
`Fibrinogen (g/1)
`
`MCM(pg)
`
`MCHC(g/dl)
`
`MCV(fl)
`
`Average
`
`SEM
`
`86.69
`88.54
`84.54
`59.85
`58.54
`57.00
`9.11
`9.92
`9.55
`31.00
`34.31
`33.73
`5.70
`6.69
`6.40
`9.08
`9.69
`9.77
`7.24
`8.59
`8.13
`4.54
`3.92
`4.27
`10.00
`10.38
`9.45
`29.31
`29.31
`28.00
`34.23
`35.23
`34.82
`
`14.02
`11.54
`12.64
`4.26
`3.46
`4.24
`1.85
`1.85
`1.39
`3.86
`5.43
`3.60
`2.05
`3.71
`2.15
`1.26
`1.35
`1.00
`1.11
`2.65
`1.75
`1.78
`1.38
`I. 71
`1.41
`1.78
`0.66
`3. 12
`3.43
`2.89
`3.49
`3.93
`4.17
`
`A
`B
`C
`A
`B
`C
`A
`B
`C
`A
`B
`C
`A
`B
`C
`A
`B
`C
`A
`B
`C
`A
`B
`C
`A
`B
`C
`A
`B
`C
`A
`B
`C
`
`n
`
`13
`13
`13
`13
`13
`11
`13
`13
`11
`13
`13
`11
`10
`11
`11
`13
`13
`11
`13
`13
`11
`13
`13
`11
`13
`13
`11
`13
`13
`11
`13
`13
`11
`
`Average
`
`SEM
`
`136.38
`142.46
`136.69
`66. 17
`64.77
`64.38
`10.75
`11.88
`12.12
`32.75
`36.00
`36.62
`4.28
`3.49
`5.78
`10.28
`10.44
`10.65
`7.18
`8.63
`7.88
`3.75
`3.62
`3.77
`10.67
`11.54
`11.38
`33.00
`33.38
`33.15
`31.92
`34.54
`34.54
`
`15.73
`18.08
`16.13
`6.05
`3.62
`3.34
`1.89
`0.89
`1.03
`4.78
`2.80
`3.08
`2.54
`2.53
`1.56
`0.96
`0.84
`0.92
`1.27
`1.94
`2.34
`1.30
`1.64
`1.25
`1.49
`1.28
`1.00
`2.52
`1.60
`1.99
`4.42
`2.65
`1.91
`
`n
`
`13
`13
`13
`12
`13
`13
`12
`13
`13
`12
`13
`13
`13
`13
`13
`12
`13
`13
`12
`13
`13
`12
`13
`13
`12
`13
`13
`12
`13
`13
`12
`13
`13
`
`Average
`
`SEM
`
`163.23
`168.00
`170.69
`64.00
`64.77
`65.08
`9.36
`10.80
`11.62
`31.38
`35.54
`37.69
`3.31
`5.38
`5.02
`9.96
`10.63
`10.77
`6.68
`7.03
`6.85
`3.38
`3.54
`3.69
`9.31
`10.15
`10.85
`30.46
`30.38
`30.92
`31.38
`34.15
`35.00
`
`16.79
`16.89
`19.62
`2.69
`4.10
`3.36
`1.52
`1.39
`1.54
`5.53
`4.50
`3.29
`2.03
`4.25
`2.35
`1.08
`1.48
`0.63
`1.43
`1.61
`1.61
`0.92
`1.55
`1.20
`1.26
`1.29
`1.29
`1.39
`1.08
`2.62
`4.32
`3.55
`2.63
`
`n
`
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`13
`
`A: control; B: 4 ml iron dextran; C: 8 ml iron dextran.
`
`34
`
`Arzneim.-Forsch./Drug Res. 41 (I), Nr. I ( 1991)
`Geisser et al. -
`Iron dextran
`
`PGR2020-00009
`Pharmacosmos A/S v. American Regent, Inc.
`Petitioner Ex. 1042 - Page 3
`
`

`

`II
`
`,.
`
`C g
`
`0
`
`5
`weeks after treatment
`Fig. 5: Development of serum iron during the first 8 weeks of the test
`period.
`
`a
`
`0
`
`5
`weeks after treatment
`Fig. 6: Development of erythrocyte number during the first 8 weeks of the
`test period.
`
`a
`
`11
`
`1 11
`0 .::. .,
`~ 0 "" :,
`
`Cl
`
`..!
`
`0
`
`5
`weeks after treatment
`Fig. 7: Development of leukocyte number during thl· !"11 , 1 ~ "~cks of the
`test period.
`
`0
`
`5
`weeks after treatment
`Fig. 8: Development of fibrinogen during the first 8 weeks of the test pe-
`riod.
`
`a
`
`-;;;
`n.
`'-'
`:I:
`t.)
`:::.
`
`11
`
`5
`weeks after treatment
`Fig. 9: Development of MCH during the first 8 weeks of the test period.
`
`0
`
`a
`
`3.3. Plasma protein
`The plasma protein concentration increases in all three
`groups within the first 5 weeks significantly (p < 0.0 I)
`and remains at this level for group 2 up to the 8th week.
`In group I it drains off up to 8 weeks, while in group 3 it
`increases once again slightly. In comparison to the initial
`values all values after 8 weeks are significantly different
`(p < 0.01). Among the groups there are only numerical
`but no significant differences.
`
`3.4. Hemoglobin
`The development of the hemoglobin concentration is in(cid:173)
`teresting. In group I and 2 the values after 5 and 8 weeks
`are significantly different (p < 0.05 resp. p < 0.01) from
`the initial values and also different (p < 0.05) from the
`values after 8 weeks. The hemoglobin concentration
`passes in all the three groups through a maximum, which
`is significant in group I and 2. In group 3 the value after
`5 weeks is significantly different (p < 0.0 I) from the in(cid:173)
`itial value, but only numerically different from the 8th
`week value. The differences among the different groups
`are as follows: After 5 weeks only the value of group 3 is
`significantly different (p < 0.05) from group I. After 8
`weeks the values of group 2 and 3 are significantly dif(cid:173)
`ferent (p < 0.05) from that of group I.
`
`3.5. Hematocrit
`The development of the hematocrit values for the groups
`1 and 2 is inconspicuous i.e. without any significance. In
`group 3 the values after 5 and 8 weeks are significantly
`different from the initial value (p < 0.05 resp. p < 0.0 I).
`Similarly the values of group 3 after 5 and 8 weeks are
`significantly different from the values of group 1 and 2
`(p < 0.05) .
`
`3.6. Iron
`The fluctuations of serum iron are considerable in gen(cid:173)
`eral. Still the development between weeks O and 8 can be
`denoted as reducing. Thereby the value of the 5th week
`of group 2 and the value of the 8th week of group I are
`significantly different (p < 0.05) from the beginning val-
`
`..
`
`,.
`
`~
`
`~
`~ JO
`t.)
`:I:
`t.)
`:::.
`
`"'
`..
`
`weeks after treatment
`Fig. 10: Development of MCHC during the first 8 weeks of the test period.
`
`..
`
`ll
`
`,.
`
`~
`
`E.
`(i
`:::.
`
`........ f
`----
`
`"'
`
`5
`weeks after treatment
`Fig. 11: I >n dupment of MCV during the first 8 weeks of the test period.
`
`0
`
`Arzneim.-Forsch./Drug Res. 41 (I), Nr. 1 ( 1991)
`Geisser et al. -
`Iron dextran
`
`35
`
`PGR2020-00009
`Pharmacosmos A/S v. American Regent, Inc.
`Petitioner Ex. 1042 - Page 4
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`

`

`ues. Similarly the value of group 2 of the 5th week is
`significantly different (p < 0.01) from the comparative
`value of group 3.
`
`3. 7. Erythrocytes
`The volume of erythrocytes develops inconspicuously.
`The only significant differences are the following: After
`5 weeks the increase in case of group 1 in comparison
`with the initial values (p < 0.05), after 8 weeks the in(cid:173)
`crease in group 3 compared with the initial values (p
`< 0.05) and the difference from group 1 (p < 0.01).
`
`3.8. Leukocytes
`The behavior of the leukocyte numbers in blood is sim(cid:173)
`ilar, i.e. decreasing little from week O to week 8. Only the
`value of group 2 at the 5th week is significantly different
`(p < 0.05) from the value at the 8th week and also from
`the value of the 5th week of group 1 (p < 0.05).
`
`3.9. Fibrinogen
`The behavior of the fibrinogen concentrations is incon(cid:173)
`spicuous with no differences among the groups. The nu(cid:173)
`merical reductions during the 8 weeks are not significant.
`
`3.10. MCH, MCHC and MCV
`MCH (mean corpuscular hemoglobin concentration in
`pg) develops for group 1 and 2 through a maximum after
`5 weeks (p < 0.05 in comparison after 8 weeks). For
`group 3 the values after 5 and 8 weeks are significantly
`(p < 0.01) higher than the starting values. The numeric
`maximum for group 3 is observed after 5 weeks. The dif(cid:173)
`ferences between the groups are not significant, except
`the value of group 3 as compared to group 2 after 8 weeks
`(p < 0.01).
`MCHC (mean corpuscular hemoglobin concentration in
`g/1) develops in case of all three groups in the same man(cid:173)
`ner. The values after 5 weeks are significantly different
`(p < 0.01) from the initial values and the values after 8
`weeks. There are no differences among the groups.
`MCV (mean corpuscular volume in fl) diminishes in
`group 1, but not significantly. In group 2 and 3 the values
`remain constant.
`
`++
`( +)
`++
`
`3.11. Histological finding of spleen
`Group 1: without iron dextran 20 %. Random sample test
`preparation of animal No. 302: practically no detectable
`iron.
`Group 2: 4 ml iron dextran 20 % per animal. Random
`sample test preparation of animal No. 4003: in general
`little, as per rule single iron pigments with the following
`distribution:
`red pulp
`white pulp
`marginal zone
`capsule and trabecula
`Group 3: 8 ml iron dextran 20 % per animal. Random
`sample test preparation of animal No. 4036: in general
`little, but in comparison to preparation No. 4003 some
`more, often in groups remaining iron pigments with the
`following distribution:
`red pulp
`white pulp
`marginal zone
`capsule and trabecula
`Check: random sample test preparation of spleen ofa i.v.
`iron-loaded rat for the control of the reliability of the
`executed Berliner blue reaction: in general very many,
`mostly in groups remaining and to the greatest extent
`confluencing iron pigments with the following distribu(cid:173)
`tion:
`
`+
`+
`++
`
`36
`
`red pulp
`white pulp
`marginal zone
`capsule and trabecula
`
`++
`+
`+++
`( +)
`
`3.12. Local reactions
`After the injection of iron dextran 20 % no local side re(cid:173)
`actions could be observed within the first three days after
`application. There was no coloration of the skin and also
`no swelling of the tissue. This is not astonishing, because
`all batches produced by Hausmann Laboratories have
`been tested for these parameters on piglets before releas(cid:173)
`ing according to the technique published by Schmitz et
`al. [8].
`
`4. Discussion
`The efficacy of oral and parenterally applicable iron prep(cid:173)
`aration on hematology, growth and the quality of the veal
`of calves has already been described by different authors
`[ 1, 2, 3, 5, 6, 9]. But the great problem lies in the fact that
`the quality of veal is judged differently from country to
`country and that consumers have different attitudes to
`this. The object of this study was to show that with a
`definite dosage the quality of veal, according to typical
`Swiss criteria, can be assessed as first class. Thereby
`mainly the criterion of color arises. The evaluation ac(cid:173)
`cording to the parameter "red" veal in the different
`groups showed the following: group I without iron dex(cid:173)
`tran 20 % showed 3 calves with "red" veal, group 2 with
`4 ml iron dextran 20 % and group 3 with 8 ml iron dex(cid:173)
`tran 20 % 5 calves each.
`It should be noted that the color cannot be brought in
`connection with haemoglobin values and other blood
`parameters. This is also a result obtained by Disler [3].
`The influence e.g. of the rate of bleeding to death is not
`to be overlooked. The histological findings are more pro(cid:173)
`nounced. In non-treated animals practically no iron was
`found in the spleen with the histology test; in case of an(cid:173)
`imals treated with 4 ml iron dextran 20 % traces of iron,
`and in those treated with 8 ml iron dextran 20% some
`more than traces of iron was detected. In both groups
`treated with iron dextran 20 % the distribution in the in(cid:173)
`dividual parts of the spleen does not show any clear dif(cid:173)
`ference. In general in the group treated with 8 ml only
`very little iron was histologically detectable.
`From the measured values of haemoglobin, MCH and
`MCHC it results that after a maximum at 5 weeks, at 8
`to 10 weeks, again a more distinctive anaemia can be
`observed. This is also shown histologically by the empty
`iron depots in the spleen in all the three groups.
`Most interesting is the increase in weight of the group
`with 8 ml iron dextran 20 % ( 1600 mg iron). The higher
`increase of an average of 6.5 kg per calf means better
`growth. The plasma protein shows the same effect.
`Whereas the plasma protein increases in group 1 only by
`4.2 g/1, the corresponding increase in group 3 is almost
`double, namely 8.1 g/1. This positively influences the
`quality of veal. Tests were not conducted in this respect.
`From numerous tests it results that addition of iron (11)(cid:173)
`salt to food leads to interactions with the food and gen(cid:173)
`erally only to a partial absorption i.e. about 20 % (de(cid:173)
`pending on the degree of anaemia and the composition
`of the food). These studies show that a single dose of 8
`ml iron dextran 20 % has a good effect on growth without
`negatively influencing the quality of veal. The least stan(cid:173)
`dardized parameter is to be searched in the absorption
`of iron from food as also with the eaten quantity of food
`from calf to calf. To achieve a more exact dosage of iron
`for all animals it would be meaningful to use food in fu(cid:173)
`ture without iron addition. By a single depot dosage of 8
`ml ( 1600 mg Fe) to 12 ml (2400 mg Fe) iron dextran 20 %
`
`Arzneim.-Forsch.1Drug Res. 41 (I), Nr. I ( 1991)
`Geisser et al. -
`Iron dextran
`
`PGR2020-00009
`Pharmacosmos A/S v. American Regent, Inc.
`Petitioner Ex. 1042 - Page 5
`
`

`

`the iron requirement, as calculated under 2.2, can be met.
`8 ml iron dextran 20 % correspond to the iron require(cid:173)
`ment only for the variant with iron fortified food. With(cid:173)
`out iron fortificated food, a minimum of I 0-12 ml iron
`dextran 20 % is necessary. This variant has two advan(cid:173)
`tages: I. better standardization of the iron supply for the
`calves and 2. purchase of cheaper food without iron,
`which is not insignificant for the economic efficiency.
`A study on full term infants, carried out by Olivares et
`al. [7], has shown that a intramuscular single dose of 150
`mg of iron as iron dextran at birth affords a nutritional
`advantage in iron status for up to 15 months, when com(cid:173)
`pared with a unfortified group. The group fed an iron
`fortified milk, containing 15 mg iron as ferrous sulfate
`and I 00 mg/I ascorbic acid, however, caught up with the
`iron dextran group by 9 months, whereby the group had
`to take it from the 3rd to the 15th month. Here not only
`problems with the compliance arise but also with all
`kinds of possible interactions of the used ferrous sulfate
`with the food stuffs. From this point of view the problems
`of oral iron absorption and the efficacy of iron dextran
`in our study are comparable with the study of Olivares.
`Both studies show that these problems can be solved with
`a single dose application of a well defined iron dextran
`complex. Nevertheless, the indication for the use of a par(cid:173)
`enteral iron dextran preparation has to be taken very
`carefully in infants.
`
`5. Literature
`[I) Boehnke, E., Gropp, J., Bayer. Landwirtsch. Jahrbuch. 56,
`571 (1979)-[2] Bilnger, U., Schmoldt, P., Steinhardt. M., Mh.
`Vet. Med. 42, 20 ( 1987) -
`[3] Disler, B., Untersuchungen zur
`Beurteilung der Eisenversorgung bei Mastklilbern nach der
`Schlachtung, Dissertation, Veterinlir-medizinische Fakultlit.
`[4] Geisser, P., Arzneim.-Forsch./
`Universitlit Bern ( 1986) -
`(5) Mollerberg, L., Ehlers, T..
`Drug Res. 40 (II), 754 (1990) -
`Jacobsson, St. 0., Johnson, S., Olsson, I., Acta vet. scand. 16,
`(6) Mollerberg, L., Ekman, L., Jacobsson, St. 0.,
`197 ( 1975) -
`Acta vet. scand. 16, 205 ( 1975) -
`(7) Olivares, M. , Chadud, P ..
`Pizarro, F. , Stekel, A., Nutr. Rep. Int. 40, 577 ( 1989) -
`[8]
`Schmitz, H., Schaub, E., Millier, A., Schweiz. Arch. Tierheilk.
`ll8, 441 (1976) -
`(9) Van Hellemond, K. K., Sprietsma. J. E ..
`Landbouwk. Tijdschr./pt 89, 184 ( 1977)
`
`Acknowledgements
`We thank Dr. Hans Lutz, Faculty member of the veterinary med(cid:173)
`ical clinic of the University of Zurich for the careful conduct of
`the analysis; Mr. Pius Millier, master of the calves, Walterschwil,
`for surrendering the calves; Mr. Ernst Forrer, Ciba-Geigy, for the
`good coordination and support in planning the test. We thank
`Mrs. Irma Thiller for typing and Mr. Silvio Ramsauer for the
`technical drawings.
`
`Correspondence: Dr. rer. nat. P. Geisser, Rechenstr. 37,
`P.O.B. 983, CH-900 I St. Gallen (Switzerland)
`
`Antidiabetika
`Antidiabetics
`
`Influence of Captopril on Glucose and Fatty Acid Oxidation
`in Human Thrombocytes and Mononuclear Leucocytes
`
`R. Haeckel und D. Colic
`
`Summary
`Captopril (CAS 625 71-86-2) may be beneficial for the
`treatment of diabetes because of its activating effect on
`
`Zentra/krankenhaus St. Jiirgen-Strasse. /11sti1111 fiir
`Laboratori11111s111edi=in, Bremen (Fed. Rep. of Germany)
`
`Arzneim.-Forsch./Drug Res. 41 (I), Nr. I ( 1991)
`Haeckel et al. - Captopril
`
`peripheral glucose consumption besides its well known
`blood pressure degradation. The glucose oxidation has
`been found to be activated by captopril in thrombocytes
`and mononuclear leucocytes, cell types ll'hich are usually
`considered to be independent from insulin.
`Because the oxidation of pynrvate labelled in position C(cid:173)
`l but not of 2- 14C-pyruvate and of l- 14C-acetate was en(cid:173)
`hanced, captopril most probably stimulated the pyrm·ate
`decarboxylation reaction. The metabolism of glucose la(cid:173)
`belled in positions 1 and 6 ll'as equally activated by cap-
`
`37
`
`PGR2020-00009
`Pharmacosmos A/S v. American Regent, Inc.
`Petitioner Ex. 1042 - Page 6
`
`