Д.А.Кузнецев. Университет им.Н.И.Пирогова(РГМУ)

A NOVEL FAMILY OF MEDICINAL NANOPARTICLES TO PROMOTE THE MAGNETIC ISOTOPE EFFECT FOR PHARMACOLOGICAL PURPOSES

Dmitry A. KuznetsovVladimir P. Chekhonin

N.N. Semenov Institute for Chemical Physics, Russian Academy of Sciences, Moscow, Russia

Department of Medicinal Nanobiotechnology, N.I. Pirogov Russian State Medical University, Moscow, Russia

THE CREATINE KINASE ACTIVE SITE NANOTOPOLOGY

1.0

25

24 26

The rate of ATP formation by mitochondria (A) and by creatine kinase (B) as a function of magnesium isotope

The yield of ATP is given in mmole/g total protein

intact mitochondria

mitochondria subjected to a selective blockade of oxidative phosphorylation by 1-methylnicotine amide.

A B

ION – RADICAL PAIRS FORMATION

(SINGLET – TRIPLET PATH SHIFT)

MECHANISM

OF THE 25Mg MAGNETIC ISOTOPE EFFECT

EXPRESSED

IN A BIOLOGICAL PHOSPHORYLATION

PRECESSES

(Mt-CK)

THE CK CATALYTIC SITE NANOTOPOLOGY IMPACT ONTO THE CK CATALYTIC SITE NANOTOPOLOGY IMPACT ONTO THE REAGENTS ELECTRON DENSITY OVERLAPTHE REAGENTS ELECTRON DENSITY OVERLAP

A combined systemic solution of both Schroedinger and Poissone equations processed

The GPK reaction ion-radical mechanism

A phosphorylation rate expressed as the yield (Y) of -[32P]ATP produced by 1 mg of pure enzyme per 1 min related to the ATP synthesis rate (Y0) directed by enzyme sample with a natural

abundance of Mg2+ isotopes as a function of 25Mg content in CK active sites.

Buckminsterfullerene(C60)-2-(butadiene-1-yl)--tetra(o--aminobutyryl-o-phtalyl)porphyrin

PORPHYLLERENE – MC16

N

N

N

N

Fe

COO- -OOC

-OOCCOO-

COO-

COO-

-OOC

-OOC

CH2

CHH2N

CH2

-OOC

H2C

HC NH2

CH2

COO-

CH2

HC NH2

CH2

COO-

CH2

CHH2N

CH2

-OOC

Mg2+

Mg2+2

2

PMC16 CATIONITE PROPERTIES AND THE NANOCLUSTERS FORMATION PMC16 CATIONITE PROPERTIES AND THE NANOCLUSTERS FORMATION AS A FUNCTION OF pHAS A FUNCTION OF pH

Blue arrow shows the iron-dextrane sphere exclusion limit

1.15nm

14.8nm

6.4nm

10.2nm

3.2nm4.7nm

pH

, portion of the total PMC16 magnesium

Blue arrow shows the iron-dextrane sphere exclusion limit

THE EFFECT OF A PMC16 – TARGETED DELIVERY OF THE EFFECT OF A PMC16 – TARGETED DELIVERY OF MgMg2+2+ ON THE DOXORUBICIN (DXR) PRE – SUPPRESSED ON THE DOXORUBICIN (DXR) PRE – SUPPRESSED ATP PRODUCTION IN RAT MYOCARDIUMATP PRODUCTION IN RAT MYOCARDIUM

0.8 DL50 DXR, i.v., 6 hrs → PMC16, i.v., 6 hrs

CK

Rela

tive A

ctiv

ity

Mg 2+InfluxFree Protons Excess

SYNERGISM OF THE MITOCHONDRIAL MATRIX CK ACTIVITY, MAGNESIUM CATIONS INFLUX AND THE

FREE PROTONS EXCESS DEGREE

The isolated rat myocardium mitochondria tested.

Yellow / Red stands for the spinless / spin Mg isotopes ratio.

SYNERGISM OF THE ATP YIELD, OXYGEN CONSUMPTION AND THE Mg2+ INFLUX IN THE PERFUSED ISOLATED

RABBIT HEART MUSCLE TISSUEA

TP y

ield

, Y/Y

o

O2

ConsumptionMg2+ Influx

ATP y

ield

, Y/Y

o

O2

ConsumptionMg2+ Influx

A B

A – Zero spin magnesium test B – Magnetic magnesium test

A

D

B

C

ELECTRON TRANSMITTING MICROPHOTOGRAMS OF THE RAT MYOCARDIOCYTIC PERINUCLEAR AREAS

A, C – PMC16 related hypoxia preventing effect

B – Inhalation oxygen deficiency hypoxia model

D – Intact myocardium

PMC16 CLUSTER POSITIONING INSIDE THE RAT MYOCARDIOCYTIC MITOCHONDRIAL MEMBRANE IN

METABOLIC ACIDOSIS (a, c) AND IN NORMAL CONDITIONS (b, d)

a, b – Laser contrast (Nanofinder-S-6A) images

C, d – Confocal scanning microscopy

PMC16 nanoclusters immobilized on acetyl cellulose membrane

a – LCM, pH 7.00b – LCM, pH 8.40c – AFM, pH 8.80

A HIGHLY SELECTIVE TRAGETING OF PMC16 NANOPARTICELS A HIGHLY SELECTIVE TRAGETING OF PMC16 NANOPARTICELS TOWARDS THE RAT HEART MUSCLE IN A COURSE OF THE LONG – TERM TOWARDS THE RAT HEART MUSCLE IN A COURSE OF THE LONG – TERM ADMINISTRATION OF AN EXTRA LOW DRUG DOSAGEADMINISTRATION OF AN EXTRA LOW DRUG DOSAGE

NOTE: DXR, 20 mg/kg/24 hrs, i.v.:

MNA, 10 mg/kg/24 hrs, i.v.:

CONCLUSIONSCONCLUSIONSPORPHYRINE ADDUCTS OF FULLERENE CPORPHYRINE ADDUCTS OF FULLERENE C6060, , A NEW FAMILY OF MEDICINAL NANOPARTICLES A NEW FAMILY OF MEDICINAL NANOPARTICLES TO MEET THE FOLLOWING EXPECTATIONS:TO MEET THE FOLLOWING EXPECTATIONS:

• Tissue-selective targeted delivery of Tissue-selective targeted delivery of 2525MgMg2+2+ paramagnetics paramagnetics to the porphyrin-signaling cell compartments in to the porphyrin-signaling cell compartments in myocardiummyocardium and, to a lesser degree, and, to a lesser degree, lymphocyteslymphocytes

• The The 2525MgMg2+2+-related magnetic isotope effect to promote a -related magnetic isotope effect to promote a local and fast, jump-up increase of ATP production, even in local and fast, jump-up increase of ATP production, even in the tissue oxygen deplete of any sortthe tissue oxygen deplete of any sort

• Smart nanoparticles behavior while in Mt-membrane, Smart nanoparticles behavior while in Mt-membrane, i.e.i.e. a a gradual gradual 2525MgMg2+ 2+ release that occurs exclusively in response release that occurs exclusively in response to the tissue hypoxia-caused metabolic acidosisto the tissue hypoxia-caused metabolic acidosis

• Low toxic, safe, efficient and completely eliminable (CLow toxic, safe, efficient and completely eliminable (C6060--clearing / porphyrin metabolizing) agents, all suitable for clearing / porphyrin metabolizing) agents, all suitable for either short or long term administration schemeseither short or long term administration schemes