Cover letter

January 08, 2020

Editorial Department of Journal of constructional steel research

University of Concepcion

Víctor Lamas 1290, Concepción, Región del Bío Bío, Chile

Dear Editor of Journal of constructional steel research

I wish to submit a manuscript for consideration of publication in Journal of constructional steel

research. The manuscript is entitled “Seismic performance evaluation of eccentrically braced

steel frames placed in Chile”.

I confirm that this work is original and has not been published elsewhere, nor is it currently

under consideration for publication elsewhere.

This paper is intended to evaluate the seismic performance of structural archetypes designed

according to Chilean code NCh433 modified 2011 after Maule earthquake and AISC 341-10

specification subjecting the structures to different nonlinear static and dynamic analysis

according FEMA P695. I believe this manuscript is appropriate to be published in this journal

because it shows the seismic performance of residential steel structures which haven’t been

widely implemented in a seismic country as Chile so far.

Thank you very much for your consideration.

Yours Sincerely,

Mr Andrés Riffo

University of Concepcion

Víctor Lamas 1290, Concepción, Región del Bío Bío, Chile

Tel: (56) 41220 4000; Fax: (56) 41222 74 55

E-mail: [email protected]

Graphical Abstract

1
 Preparation of nimodipine microspheres contained in a solid
dispersion by the emulsion-solvent-diffusion method and evaluation
of their in vitro and in vivo characteristics

Hua Zhanga,*, Xu Wanga, Min Yanga, Y. Kawashb

a
Shenyang Pharmaceutical University, No.103, Wenhua Road, Shenyang 110016, China
b
Gifu Pharmaceutical University, 5-6-1 Mitahora-higashi, Gifu 502, Japan

Raspberry-like assembly of nanogels was prepared by Michael addition of acrylate-modified

cholesterol-bearing pullulan nanogel and thiol-modified poly (ethylene glycol) cross-linker.

Interleukin 12, an immunostimulatory cytokine, is encapsulated and released over a

prolonged period of time both in vitro and in vivo.

Title page

2
Preparation of nimodipine microspheres contained in a solid dispersion by
the emulsion-solvent-diffusion method and evaluation of their in vitro and
in vivo characteristics

Hua Zhanga,*, Xu Wanga, Min Yanga, Y. Kawashb

a
Shenyang Pharmaceutical University, No.103, Wenhua Road, Shenyang 110016, China
b
Gifu Pharmaceutical University, 5-6-1 Mitahora-higashi, Gifu 502, Japan

Corresponding author:
Corresponding author: Hua Zhang*

Mailing address: Shenyang Pharmaceutical University, No.103, Wenhua Road, Shenyang

110016, China

Tel.: +86-24-23986353; Fax: +86-24-23986355

Mobile phone: 13………..

E-mail: [email protected]

Abstract:

To prepare nimodipine immediate and sustained release microspheres in a solid

dispersion by the emulsion-solvent-diffusion method in a single step. Special attention was

devoted to improving the dissolution rate of the drug and then the controlled release of drug

from the microspheres. Eu E100 and Aerosil were employed as a solid dispersion base for the

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immediate release microspheres, while HP55 and aerosil were employed as a solid dispersion

base for the sustained release microspheres, while ethyl cellulose (EC) was selected as a

retarding agent to control the drug release rate. Micromeritic properties of the microspheres,

the drug dispersion state and the drug release profiles of the microspheres were investigated.

Relative bioavailabilities of two types of microspheres were compared with Nimotop TM, a

commercial nimodipine tablet, in a study in 3 healthy dogs. Two types of microspheres i.e.

immediate release and sustained release microspheres could be successfully prepared through

adjustment of the ratio of nimodipine to the functional additives. The particle size of the

resultant microspheres was mainly controlled by the agitation speed during the manufacturing

process. Following X-ray, DTA and SEM analysis, it was found that nimodipine was highly

dispersed and present in the microspheres in an amorphous state. The drug release rate from

the microspheres was very dependent on the type and amount of the functional additives in

the formulation. The relative bioavailabilities of the immediate release microspheres and

sustained release microspheres, compared with the NimotopTM tablet, were 118.2% and

96.1%, respectively. The emulsion-solvent-diffusion method is a simple and efficient

technique for designing functional microspheres for poorly water-soluble drugs.

Keywords: Nimodipine; Solid dispersion; Microsphere; Sustained release

1. Introduction

Solid dispersion is one of the most efficient techniques to improve the dissolution rate of

poorly water-soluble drugs, leading to an improvement in the relative bioavailability of their

formulations. At present, the solvent method and the melting method are widely used in the

preparation of solid dispersions. In general, subsequent grinding, sieving, mixing and

granulation are necessary to produce the different desired formulations.

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 The spherical crystallization technique is a novel agglomeration method performed in a

liquid system which was developed by Kawashima in the 1980s [1]. In this system,

agglomeration and spheroidization can be carried out simultaneously during the

crystallization process of a substance with a bridging liquid by means of stirring. In the

beginning, the spherical crystallization technique was mainly used in direct tabletting

technology, because crystallization and agglomeration could be carried out simultaneously in

a single step. The resultant agglomerates produced exhibited dramatically improved

flowability, packability and compressibility [2, 3]. Later on, functional drug devices such as

microspheres [4], microcapsules [5], microballoons [6], and biodegradable nanospheres [7]

were developed using the emulsion-solvent-diffusion method, one of the spherical

crystallization techniques involving the introduction of a functional polymer into the system.

In previous studies we have designed sustained release mirospheres for a water soluble drug

[8], a water insoluble drug [9] and an oily drug [10] using this technique.

2. Materials and methods

2.1. Materials

Nimodipine (Shandong Xinhua Pharmaceutical Co. Ltd. China) was used as a model

drug ， Eudragit E (Eu E100, Röhm Pharma, Germany) and hydroxypropylmethylcellulose

phthalate (HPMCP, Shinetsu Chemical Co. Ltd. Japan) were selected as a dispersion base,

aerosol (pass 400 mesh, Qingdao Ocean Chemical Co. Ltd. China) as a absorbent was added

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to promote drug dispersion and increase the mass for compact consolidation of the resultant

microspheres. Ethylcellulose (EC 10 cp, Shanghai Colorcon Co. Ltd. China) was used as a

retarding agent to control the drug release. Acetone was selected as a good solvent, distilled

water was used as a poor solvent, and dichloromethane (Shenyang Chemical Co. Ltd.) was

chosen as a bridging agent due to its good wettability with regard to both the drug and

polymers, and its immisciblity with the poor solvent. A small amount of surfactant, such as

poloxamer (F188, Shenyang Pharmaceutical Factory, China), was added to the poor solvent

in order prevent sticking to the baffle or the agitator. NimotopTM (Bayer com., Germany) was

selected as a reference formulation for the relative bioavailability test. All solvents used were

of analytical grade.

2.2. Preparation of immediate release microspheres of nomodipine

Nimodipine (0.6 g) and Eu E100 (1.2, 2.4 and 3.6 g) were dissolved in a mixture of good

solvent (acetone, 10 ml) and bridging liquid (dichloromethane, 2.5 ml), and the dispersion

agent (aerosol, 1.2, 2.4 and 3.6 ml) was added to the drug-polymer solution system. The poor

solvent (distilled water, 150 ml) containing poloxamer (4%, 3 ml) was placed in a 500 ml

flask fitted with three baffles to improve agitation during the agglomeration process. When

the drug-polymer solution was added to the poor solvent at a temperature of 25C and 400–

600 r/min stirring with a propeller type agitator, the drug-polymer solution immediately

dispersed as droplets to form a quasi o/w emulsion, and the emulsion droplets gradually

solidified along with the diffusion of the good solvent from the droplets into the poor solvent.

Finally, the coprecipitated microspheres of the drug-polymer were filtered and dried in oven

at 50C for more than 6 h to reduce the residual solvent in accord with the tolerance given in

Ch.P.

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2.3. Preparation of sustained release microspheres of nimodipine

In the preparation of the sustained release microspheres, the process and the equipment was

the same as that used for the immediate release microspheres. The formulations were as

follows: Nimodipine: HP55: aerosol: EC = 1: 2: 7: (0.5, 0.625 and 0.75) were dissolved in the

mixture of good solvent (acetone, 11 ml) and bridging liquid (dichloromethane, 7 ml), then

the drug-polymer solution was poured into the poor solvent (distilled water, 200 ml) with the

temperature controlled at 15–20 C under stirring at 700 r/min. After agitating the system for

20–40 min, the emulsion droplets gradually solidified through coacervated droplets, and the

resulting microspheres were filtered and dried using the same method as for the immediate

release microspheres.

3. Results and discussion

3.1. Preparation mechanism

In this study, acetone and dichloromethane, in which nimodipine is readily soluble, were

used as a good solvent and bridge liquid to dissolve the drug and pH- dependent polymers,

and distilled water containing 4% (w/v) poloxamer was used as a poor solvent to precipitate

both nimodipine and the pH-dependent polymers. Gradually, the emulsion droplets solidified

along with diffusion of the good solvent, e.g., drug and polymers were coprecipitated in the

droplets, leading to the formation of microspheres, i.e., dichloromethane as the bridging

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liquid was commixed with the good solvent, and when the good solvent in the droplets

diffused into the poor solvent, the residual dichloromethane in the droplets bridged the

aerosil, coprecipitated drug and polymer to form agglomerates. The aerosil acts as a

dispersing agent and mass compactor, because coacervation droplets formed from the drug-

polymer droplets during the solidifying period were very sticky and readily coalesced, while

the introduction of Aerosil efficiently prevented coalescence and produced compact

microspheres. In addition, the aerosol also dramatically improved the drug dispersion,

resulting in the formation of a solid dispersion.

3.2. Micromeritic properties of the microspheres

As discussed above, the mean particle size (D50) or size distribution of the microspheres

was mainly controlled by the agitation speed. The average diameter of the microspheres was

controlled mainly by the agitation speed during the formation of quasi-emulsion droplets

during the initial stage. Fig. 2 shows the change in the D50 of the microspheres with regard to

the stirring speed, i.e., increasing the agitation speed reduced the particle size. It was found

that the temperature of the solvent system also affected the properties of the resulting

microspheres. Increasing the temperature of the liquid system resulted in a reduction in

particle size, and tended increase the sphericity. In this study, under the optimum conditions

for producing immediate release microspheres, which were determined by the orthogonal

design method, the total recovery was 97%. The micromeritic properties of the resultant

microspheres are shown in Table 1.

4. Conclusion

By combination of a polymeric crystallization technique and a solid dispersion

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technique, the immediate and sustained release microspheres could be successfully produced

in a solid dispersion state. Aerosil as a drug dispersion agent and fillers helped prevent the

coalescence of sticky polymer droplets during the preparation process, the method was highly

reproducible and offered a high recovery.

Declaration of interest

The authors report no conflicts of interest. The authors alone are responsible for the

content and writing of this article.

Acknowledgements

The authors acknowledge the financial support received from Council for Technical

Education (CTE), for their support and encouragement in carrying out his college work.

References

[1] Francis MF, Lavoie L, Winnik FM, Leroux JC. Solubilization of cyclosporine A in

dextran-g-polyethyleneglycolalkylether polymeric micelles. Eur J Pharm Biopharm

2003;56(3):337-46.

[2] Joshi R, Feldmann V, Koestner W, Detje C, Gottschalk S, Mayer HA, et al.

Multifunctional silica nanoparticles for optical and magnetic resonance imaging. Biol

Chem 2013;394(8):125-35.

Figure and Table legends

Fig. 1. Effect of tablet geometry on drug release.

Fig. 2. Pathological features of specimens of the: (A) anterior mediastinum and (B) hepatic

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tumors demonstrate similar morphology as described in the text. (C, D) he key stains for CD5

were all positive.

Table 1 Micromeritic properties of the immediate release microspheres.

Table 2 Pharmacokinetic parameters of nimodipine after oral administrations in dogs.

Figures:

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 Fig. 1. Effect of tablet geometry on drug release.

600 g/g control

Control

A B

0.2 mm 0.2 mm

11

C D
5 m 5 m
Fig. 2. Pathological features of specimens of the: (A) anterior mediastinum and (B) hepatic

tumors demonstrate similar morphology as described in the text. (C, D) he key stains for CD5

were all positive.

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Figure samples

13
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Tables:

Table 1 Pharmacokinetic parameters of nimodipine after oral administrations in dogs

tpeak Cmax AUC0→∞ ke ka

Test preparations
(h) (ng/ml) (ng/ml·h) (h-1) (h-1)
NimotopTM 0.58 80.29 234.70 0.376 14.832
Immediate release 0.59 70.02 277.33 0.302 5.058
Sustained release 3.74 28.92 225.53 0.284 0.442

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